"Cytokine Storm" in the Phase I Trial of Monoclonal Antibody TGN1412: Better Understanding the Causes to Improve PreClinical Testing of Immunotherapeutics
Open accessAbstract
The CD28-specific mAb TGN1412 rapidly caused a life-threatening "cytokine storm" in all six healthy volunteers in the Phase I clinical trial of this superagonist, signaling a failure of preclinical safety testing. We report novel in vitro procedures in which TGN1412, immobilized in various ways, is presented to human white blood cells in a manner that stimulates the striking release of cytokines and profound lymphocyte proliferation that occurred in vivo in humans. The novel procedures would have predicted the toxicity of this superagonist and are now being applied to emerging immunotherapeutics and to other therapeutics that have the potential to act upon the immune system. Data from these novel procedures, along with data from in vitro and in vivo studies in nonhuman primates, suggest that the dose of TGN1412 given to human volunteers was close to the maximum immunostimulatory dose and that TGN1412 is not a superagonist in nonhuman primates.
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- Findingown data
TGN1412 captured in aqueous phase by immobilized anti-human Fc Ab produced greater human lymphocyte proliferative responses than air-dried TGN1412.
“Even greater human lymphocyte proliferative responses were achieved with TGN1412 captured in aqueous phase by immobilized anti-human Fc Ab”
Quote in the paperSubstantial claim - Findingown data
Human lymphocytes stimulated with aqueous TGN1412 captured by immobilized anti-Fc antibody produced greater IL-2 than with immobilized TGN1412 alone.
“In contrast, human lymphocytes stimulated with aqueous TGN1412 captured by immobilized anti-Fc Ab stimulated greater IL-2 production than observed with immobilized TGN1412 alone (D).”
Quote in the paperSubstantial claim - Findingown data
TGN1412 air-dried onto wells failed to stimulate significant production of TNF-alpha, IL-6, and IL-8 from Cynomolgus macaque diluted blood.
“In marked contrast, TGN1412 air dried onto the walls of wells failed to stimulate significant production of TNF-␣, IL-6, and IL-8 from the diluted blood (20% v/v) of Cynomolgus macaques”
Quote in the paperSubstantial claim - Findingown data
At 0.4 g/well TGN1412, approximately equivalent to the human clinical trial dose, a strong proliferative response of human lymphocytes was observed.
“In contrast, with 0.4 g/well TGN1412, approximately equivalent to the human dose administered in the clinical trial, a strong proliferative response of human lymphocytes was observed.”
Quote in the paperSubstantial claim - Findingown data
The optimal proliferative dose response of human lymphocytes to immobilized TGN1412 was between 0.4 and 2 g/well.
“Measurement of proliferative responses of human lymphocytes to immobilized TGN1412 identified the optimal dose response as between 0.4 and 2 g/well (Fig. 7A).”
Quote in the paperSubstantial claim
- Findingown data
Immobilized TGN1412 stimulated IFN-gamma production by human CD4+ lymphocytes.
“Immobilized TGN1412 also stimulated IFN-␥ production by CD4 ϩ human lymphocytes.”
Quote in the paperSubstantial claim - Findingown data
All IL-2+ human lymphocytes following immobilized TGN1412 stimulation are also positive for TNF-alpha.
“In subsequent studies, we have confirmed that all of these IL-2 ϩ human lymphocytes are also positive for TNF-␣, following stimulation with immobilized TGN1412.”
Quote in the paperNot a substantial claim - Findingown data
ELISA of 3-day culture supernatants showed high IL-2 and lower IFN-gamma levels in response to TGN1412 at low (0.4-2 g/well) but not high (20 and 200 g/well) concentrations.
“ELISA data for supernatants from 3-day cultures of human lymphocytes stimulated with immobilized TGN1412 confirmed high levels of IL-2 and lower levels of IFN-␥ in response to TGN1412 at low (0.4 to 2 g/well) but not high (20 and 200 g/well) concentrations of TGN1412.”
Quote in the paperSubstantial claim - Findingown data
Higher doses of 20 or 200 g/well TGN1412, approximately equivalent to 5 and 50 mg/kg tested in Cynomolgus macaques, produced only weak proliferative responses of human lymphocytes.
“With higher doses of 20 or 200 g/well, approximately equivalent to the doses (5 and 50 mg/kg) previously tested in Cynomolgus macaques, only weak proliferative response of human lymphocytes were observed.”
Quote in the paperSubstantial claim - Findingown data
IL-2 alone did not stimulate proliferation of human or Cynomolgus macaque PBMC.
“IL-2 alone did not stimulate proliferation of human or Cynomolgus macaque PBMC.”
Quote in the paperSubstantial claim - Findingown data
Immobilized TGN1412 increased CD25 expression on macaque CD4+ T cells relative to controls.
“Stimulation of macaque lymphocytes with immobilized TGN1412 similarly induced significant up-regulation of the activation marker CD25 on CD4 ϩ T cells compared with controls (Fig. 5, C and D), but in contrast to human lymphocytes this was not accompanied by proliferation.”
- Also stated in: § Effect of TGN1412 in Cynomolgus macaques · boosted
“Compared with negative control (C), immobilized TGN1412 (D) strongly up-regulates expression of the activation marker CD25 (IL-2R␣) on macaque CD4 ϩ lymphocytes even though they do not undergo proliferation unless IL-2 is added to the cultures.”
Quote in the paperSubstantial claim - Also stated in: § Effect of TGN1412 in Cynomolgus macaques · boosted
- Findingown data
TGN1412 captured in aqueous solution by immobilized anti-human Fc antibody produced higher levels of IL-2+ human lymphocytes than controls.
“Even higher levels of IL-2 ϩ human lymphocytes were obtained with TGN142 captured in aqueous solution by immobilized anti-human Fc Ab compared with controls (Fig. 5, C and D)”
Quote in the paperSubstantial claim - Findingown data
Human lymphocytes stimulated with a combination of aqueous TGN1412 and aqueous anti-Fc antibodies did not produce IL-2.
“Human lymphocytes stimulated with a combination of aqueous TGN1412 and aqueous anti-Fc Abs did not produce IL-2 (C).”
Quote in the paperSubstantial claim - Findingown data
The proportion of IL-2+ lymphocytes after immobilized TGN1412 was equivalent to that seen with combined PMA and ionomycin.
“The level of IL-2 ϩ lymphocytes was equivalent to the response seen with the combined stimulation of PMA and ionomycin.”
Quote in the paperSubstantial claim - Findingown data
Immobilized TGN1412 stimulated proliferative responses in up to 35% of human lymphocytes.
“Analysis of lymphocyte proliferation, quantified as precursor frequency of responding cells per 10 6 lymphocytes (Fig. 4, C and D), showed that immobilized TGN1412 stimulated proliferative responses in up to 35% of lymphocytes.”
Quote in the paperSubstantial claim - Findingown data
Cynomolgus macaque lymphocytes showed very little proliferative response to immobilized TGN1412.
“In stark contrast, Cynomolgus macaque lymphocytes showed very little proliferative response to immobilized TGN1412 (Fig. 5A)”
Quote in the paperSubstantial claim - Findingown data
TGN1412 in aqueous solution did not stimulate release of TNF-alpha, IL-6, and IL-8 from human PBMC above basal or negative control levels.
“TGN1412 (0.5-250 g/well ϭ 2.0 -1000 g/ml) added in aqueous solution and cultured for 16 -24 h with human PBMC (50,000 -200,000 PBMC/well) or human whole blood (20% v/v final concentration) did not stimulate the release of TNF-␣, IL-6, and IL-8 above basal values and responses to negative controls.”
Quote in the paperSubstantial claim - Findingown data
TGN1412 in aqueous solution, wet coated, or cross-linked with anti-human Fc Ab did not stimulate significant proliferation of human lymphocytes.
“In contrast, TGN1412 in aqueous solution, wet coated onto plates or cross-linked in aqueous solution with an anti-human Fc Ab, did not stimulate any significant proliferation of human lymphocytes.”
Quote in the paperSubstantial claim - Findingcitation
The TGN1412 used in the trial was free of nonendotoxin pyrogenic contaminants.
“Additional testing revealed the TGN1412 to be free of”
Quote in the paperSubstantial claim0 of 3 sources checked- cites [3] 2003
Not checked in the free run.
- cites [4] 2004
Not checked in the free run.
- cites [5] 2005
Not checked in the free run.
- Findingown data
Anti-CD3 and anti-CD28 costimulation strongly stimulate both human and Cynomolgus macaque lymphocytes.
“but is not supported by our finding that anti-CD3 and anti-CD28 costimulation strongly stimulate both human and Cynomolgus macaque lymphocytes.”
Quote in the paperSubstantial claim - Findingown data
Macaque lymphocytes were activated by immobilized TGN1412, as shown by up-regulation of IL-2R expression and blast transformation.
“However, macaque lymphocytes were activated by immobilized TGN1412 (as shown by the up-regulation of IL-2R expression and blast transformation), confirming binding to and interaction with macaque CD28.”
Quote in the paperSubstantial claim - Findingown data
TGN1412 evokes cytokine release and lymphocyte proliferation in vitro only when presented to PBMC in the effective manner identified in these studies.
“Second, we show that TGN1412 evokes cytokine release and lymphocyte proliferation in vitro only when presented to PBMC in the effective manner identified in these studies”
Quote in the paperSubstantial claim - Findingown data
The IFN-gamma response to immobilized TGN1412 was much smaller than that to PMA and ionomycin, which stimulated about 5- to 10-fold more CD8+ lymphocytes to produce IFN-gamma.
“However, the IFN-␥ response was overall much smaller than that observed with the mitogens PMA and ionomycin, which stimulated ϳ5to 10-fold more CD8 ϩ lymphocytes to produce this cytokine.”
Quote in the paperNot a substantial claim - Findingown data
A 6-hour stimulation of human lymphocytes with immobilized TGN1412 caused significant IL-2 production by CD4+ lymphocytes compared with controls.
“Stimulation of human lymphocytes with immobilized TGN1412 for 6 h resulted in significant production of IL-2 by CD4 ϩ lymphocytes compared with controls (Fig. 6, A and B).”
Quote in the paperSubstantial claim - Findingown data
No adverse reactions occurred in Cynomolgus macaques after administration of syringe-derived material at the human dose (0.1 mg/kg).
“Also, there were no adverse reactions in Cynomolgus macaques following administration of syringe-derived material at the human dose (0.1 mg/kg): the”
Quote in the paperSubstantial claim - Findingown data
In vitro proliferative responses to TGN1412 exhibited a bell-shaped dose-response curve.
“Our in vitro proliferative responses to TGN1412 exhibited a bell-shaped dose-response curve, with a maximum response to a concentration of TGN1412 that we calculate to be approximately equivalent to the dose of 0.1 mg/kg given to the human volunteers.”
Quote in the paperSubstantial claim - Findingown data
The level of proliferation induced by immobilized TGN1412 was equivalent to that observed with positive controls PHA and mitogenic anti-CD3 Ab.
“The level of proliferation induced by immobilized TGN1412 was profound and equivalent to the responses observed with the stimuli used as positive controls: PHA and mitogenic anti-CD3 Ab alone and in combination with conventional anti-CD28.”
Quote in the paperSubstantial claim - Findingown data
All PHA-stimulated human lymphocyte day 3 culture supernatants were positive for IL-2 and IFN-gamma by ELISA.
“In con-trast, all PHA-stimulated human lymphocyte day 3 culture supernatants were positive for IL-2 and IFN-␥ by ELISA.”
Quote in the paperNot a substantial claim - Findingown data
The optimal dose response for IL-2 production by human lymphocytes to immobilized TGN1412 was between 0.4 and 2 g/well.
“For IL-2 production measured by intracellular cytokine staining, the optimal dose response of human lymphocytes to immobilized TGN1412 was between 0.4 and 2 g/well (Fig. 7B).”
Quote in the paperSubstantial claim - Interpretationown data
The striking difference between humans and Cynomolgus macaques in in vivo toxicity of TGN1412 is believed to be a consequence of the very different way white blood cells from humans and macaques respond to TGN1412.
“The striking difference between human and Cynomolgus macaques in terms of the in vivo toxicity of TGN1412 is, we believe, a consequence of the very different way in which white blood cells from humans and macaques respond to TGN1412.”
Quote in the paperSubstantial claim - Interpretationreasoning§ "Cytokine Storm" in the Phase I Trial of Monoclonal Antibody TGN1412: Better Understanding the Causes to Improve PreClinical Testing of Immunotherapeutics
The adverse events in the Phase I trial signaled a failure of preclinical safety testing.
“signaling a failure of preclinical safety testing.”
Quote in the paperSubstantial claim - Interpretationown data
White blood cells from Cynomolgus macaques do not respond to TGN1412 in the same way as human white blood cells, whether stimulated in vitro with immobilized TGN1412 or in vivo.
“Third, we show that white blood cells from Cynomolgus macaques do not respond to TGN1412 in the same way as human white blood cells, whether the cells are stimulated in vitro, using immobilized TGN1412, or in vivo.”
Quote in the paperSubstantial claim - Interpretationown data§ "Cytokine Storm" in the Phase I Trial of Monoclonal Antibody TGN1412: Better Understanding the Causes to Improve PreClinical Testing of Immunotherapeutics
The novel in vitro procedures would have predicted the toxicity of TGN1412.
“The novel procedures would have predicted the toxicity of this superagonist”
Quote in the paperSubstantial claim - Interpretationown data
The in vitro protocols used before this study failed to predict TGN1412 toxicity in vivo because they did not present the antibody to white blood cells in a manner mimicking its in vivo presentation.
“In answer to this question, our study first provides convincing evidence that the in vitro protocols used before the present study failed to predict toxicity in vivo because they did not present the Ab to white blood cells in a manner that mimicked its presentation in vivo.”
Quote in the paperSubstantial claim- Rests on an assumption: The in vitro presentation of TGN1412 to PBMC that elicits cytokine release and proliferation mimics the in vivo presentation that caused the clinical toxicity.
- Interpretationown data
The maximum proliferative response concentration of TGN1412 in vitro is approximately equivalent to the 0.1 mg/kg dose given to human volunteers.
“Our in vitro proliferative responses to TGN1412 exhibited a bell-shaped dose-response curve, with a maximum response to a concentration of TGN1412 that we calculate to be approximately equivalent to the dose of 0.1 mg/kg given to the human volunteers.”
Quote in the paperSubstantial claim- Rests on an assumption: The in vitro concentration of TGN1412 can be converted to an equivalent in vivo dose in humans.
- Premisecitation
The TGN1412 used in the trial was not contaminated with endotoxin.
“the TGN1412 used in the trial was found not to be contaminated with endotoxin in the highly sensitive bacterial endotoxins (Limulus) test (2).”
Quote in the paperSubstantial claim0 of 1 sources checked- cites [2] 2005
Not checked in the free run.
- Interpretationown data§ "Cytokine Storm" in the Phase I Trial of Monoclonal Antibody TGN1412: Better Understanding the Causes to Improve PreClinical Testing of Immunotherapeutics
The dose of TGN1412 given to human volunteers was close to the maximum immunostimulatory dose.
“Data from these novel procedures, along with data from in vitro and in vivo studies in nonhuman primates, suggest that the dose of TGN1412 given to human volunteers was close to the maximum immunostimulatory dose”
Quote in the paperSubstantial claim - Interpretationown data
The responses measured with air-dried immobilized TGN1412 may underestimate the maximum achievable response.
“suggesting that the responses measured with air-dried (immobilized directly onto the plate) TGN1412 may be an underestimation of the magnitude of the maximum achievable response.”
Quote in the paperSubstantial claim - Premisecitation
All six recipients of TGN1412 in the 2006 clinical trial suffered life-threatening severe adverse events as a consequence of a cytokine storm.
“In 2006, these statements were contradicted when all six recipients of the CD28-specific mAb TGN1412 suffered life-threatening severe adverse events as a consequence of just such a cytokine storm (1).”
Quote in the paperSubstantial claim0 of 1 sources checked- cites [1] 2006
Not checked in the free run.
- Interpretationown data
In vitro data for Cynomolgus macaque white blood cells are entirely consistent with data from the new in vivo study in which the original preclinical study in Cynomolgus macaques was repeated and extended.
“Our in vitro data for Cynomolgus macaque white blood cells are entirely consistent with the data obtained in our new in vivo study in which the original in vivo preclinical study in Cynomolgus macaques that failed to predict the toxicity of TGN1412 was repeated and extended.”
Quote in the paperSubstantial claim - Interpretationown data§ "Cytokine Storm" in the Phase I Trial of Monoclonal Antibody TGN1412: Better Understanding the Causes to Improve PreClinical Testing of Immunotherapeutics
TGN1412 is not a superagonist in nonhuman primates.
“and that TGN1412 is not a superagonist in nonhuman primates.”
Quote in the paperSubstantial claim - Interpretationown data
In contrast to humans, TGN1412 is not a superagonist in Cynomolgus macaques.
“In summary, our data indicate that, in contrast to humans, TGN1412 is not a superagonist in Cynomolgus macaques”
Quote in the paperSubstantial claim
- May combine two claims. 15
- Not yet classified. 15
- The quote only partly states this claim. 2
- Classified as probably not a substantial claim. 2
May combine two claims.
Findingown dataImmobilized TGN1412 caused human lymphocytes to up-regulate CD25 and to proliferate.
“Stimulation of human lymphocytes with immobilized TGN1412 was accompanied by significant up-regulation of the activation marker CD25 (IL-2R␣) and proliferation.”
- Also stated in: § Effect of TGN1412 in Cynomolgus macaques · boosted
“A strong human lymphocyte proliferative response to immobilized TGN1412 was evident compared with immobilized isotype control Ab.”
Quote in the paperSubstantial claim- Also stated in: § Effect of TGN1412 in Cynomolgus macaques · boosted
Not yet classified.
FindingTGN1412 air-dried onto wells stimulated release of TNF-alpha, IL-6, and IL-8 from human PBMC.
“TGN1412 (1.0 and 10 g/well, but not 0.1 g/well) air dried onto the walls of wells in which PBMC were then cultured for 16 -24 h stimulated the release of TNF-␣, IL-6, and, especially, IL-8”
Quote in the paperNot yet classifiedMay combine two claims.
Findingown dataVery low levels of intracellular IL-2+ cells were detected in Cynomolgus macaque lymphocytes stimulated with immobilized TGN1412 at 1 g/well for 6 h, and day 3 culture supernatants were negative for IL-2 by ELISA.
“Although very low levels of intracellular IL-2 ϩ cells were detected in Cynomolgus macaque lymphocytes stimulated with immobilized TGN1412 (at 1 g/well) for 6 h, day 3 culture supernatants were negative for IL-2 measured by ELISA.”
Quote in the paperSubstantial claimMay combine two claims.
Findingown dataMacaque lymphocytes did not proliferate when exposed to immobilized TGN1412 alone; added IL-2 or rhesus CD3 antibody was required.
“No macaque lymphocyte proliferative response to immobilized TGN1412 alone was observed unless IL-2 was added to the wells.”
- Also stated in: § Discussion · plain
“in vitro Cynomolgus macaque lymphocytes did not proliferate when stimulated with immobilized TGN1412 unless IL-2 or rhesus CD3 Ab was added to cultures.”
Quote in the paperSubstantial claim- Also stated in: § Discussion · plain
Not yet classified.
FindingAddition of IL-2 to human lymphocytes stimulated with immobilized TGN1412 did not increase proliferative response.
“Addition of IL-2 to human lymphocytes stimulated with immobilized TGN1412 did not increase the proliferative response”
Quote in the paperNot yet classifiedNot yet classified.
FindingTGN1412 injected into Cynomolgus macaques at 5 and 50 mg/kg (doses used in preclinical testing) and at 0.1 and 0.5 mg/kg was well tolerated.
“TGN1412 injected into Cynomolgus macaques at the doses (5 and 50 mg/kg) used in preclinical testing and also at 0.1 and 0.5 mg/kg was well tolerated in these nonhuman primates.”
Quote in the paperNot yet classifiedNot yet classified.
FindingThe TGN1412 used in the trial passed the pharmacopoeial test for abnormal toxicity.
“Also, the TGN1412 passed the pharmacopoeial test for abnormal toxicity using mice and guinea pigs (6).”
Quote in the paperNot yet classified0 of 1 sources checked- cites [6] 2005
Not checked in the free run.
Not yet classified.
FindingWhen immobilized by drying, captured by anti-Fc antibody, or presented via an endothelial cell monolayer, TGN1412 stimulated substantial release of TNF-alpha, IL-2, IL-6, IL-8, and IFN-gamma and profound proliferation of human CD4+ lymphocytes in vitro.
“With regard to the second finding, when immobilized by drying (with warm air) onto wells of 96-well plates, captured by immobilized anti-Fc Ab, or presented via a mechanism involving the participation of an endothelial cell monolayer, TGN1412 stimulated substantial release of cytokines, including TNF-␣, IL-2, IL-6, IL-8, and IFN-␥, and evoked a profound proliferation of human CD4 ϩ lymphocytes in”
Quote in the paperNot yet classifiedNot yet classified.
FindingCynomolgus macaque lymphocytes proliferated in response to immobilized TGN1412 when recombinant human IL-2 was added.
“although they did proliferate in response to immobilized TGN1412 when recombinant human IL-2 (4 IU/well) was added to the cultures (Fig. 5B)”
Quote in the paperNot yet classifiedNot yet classified.
FindingImmobilized TGN1412 stimulated pronounced proliferation of human lymphocytes.
“Pronounced proliferation of human lymphocytes incubated with immobilized TGN1412 was clearly evident compared with immobilized isotype-matched control Ab”
Quote in the paperNot yet classifiedMay combine two claims.
Findingown dataRhesus CD3 Ab in combination with immobilized TGN1412, but not alone, stimulated much stronger proliferative responses in Cynomolgus macaque lymphocytes, equivalent to mitogen responses.
“In subsequent studies, rhesus CD3 Ab in combination with immobilized TGN1412, but not alone, stimulated much stronger proliferative responses in Cynomolgus macaque lymphocytes, responses that were equivalent to the responses to mitogen.”
Quote in the paperSubstantial claimMay combine two claims.
Findingown dataThe TGN1412 used in the trial complied with its specification and was of clinical grade.
“This analysis, using a broad range of analytical techniques, showed that the TGN1412 complied with its specification and was of clinical grade.”
Quote in the paperSubstantial claimNot yet classified.
FindingThe peak dose response for human lymphocytes was between 0.4 and 2 g/well (2-10 g/ml).
“Peak dose response for human lymphocytes was between 0.4 and 2 g/well (2-10 g/ml).”
Quote in the paperNot yet classifiedMay combine two claims.
Findingown dataNo lesions correlating with the severe complications seen in the clinical trial volunteers were detected in Cynomolgus macaques; myocarditis in one control animal and variable cardiac hypertrophy in some others were observed.
“Myocarditis was observed in one animal in the control group and variable cardiac hypertrophy in some others, but no lesions that would correlate with the severe complications seen in the volunteers at the clinical trial were detected.”
Quote in the paperSubstantial claimMay combine two claims.
Findingown dataA precursor frequency of 349,586 per 10^6 human lymphocytes was obtained with TGN1412, compared with 143 with isotype control antibody.
“A precursor frequency of 143 was obtained with isotype control Ab vs a precursor frequency of 349,586 per 10 6 human lymphocytes with TGN1412.”
Quote in the paperSubstantial claimThe quote only partly states this claim.
Findingown dataTGN1412 in aqueous solution added to a coculture of human PBMC and endothelial cells stimulated release of TNF-alpha, IL-6, and IL-8.
“TGN1412 (0.1, 1.0, and 10 g/well) added in aqueous solution to a coculture of PBMC and human endothelial cells and cultured for 24 h stimulated the release of TNF-␣”
Quote in the paperSubstantial claimNot yet classified.
FindingCynomolgus macaque blood responded to positive controls LPS and PHA despite failing to respond to TGN1412.
“Despite its failure to respond to TGN1412, blood from Cynomolgus macaques responded to the positive controls of solutions of bacterial endotoxin (LPS, 1 endotoxin unit ϭ 1 IU ϭ 100 pg/ml) and PHA (90 g/ml).”
Quote in the paperNot yet classifiedMay combine two claims.
Findingown dataCD4+ T cells were the principal responding population among proliferating lymphocytes, comprising about 85% of proliferating cells.
“Phenotypic analysis of proliferating lymphocytes identified CD4 ϩ T cells as the principle responding population (ϳ85% of proliferating cells)”
Quote in the paperNot a substantial claimMay combine two claims.
FindingcitationIn earlier in vitro studies and in the studies reported here, adding TGN1412 directly to white blood cells or cross-linking it via its Fc region in aqueous solution failed to stimulate cytokine release and cell proliferation.
“In the earlier in vitro studies, i.e., before the clinical trial, TGN1412 was simply added to white blood cells or cross-linked via its Fc region in aqueous solution, protocols that failed to stimulate release of cytokines and cell proliferation in the preclinical testing (10) and in the studies reported here.”
Quote in the paperSubstantial claim0 of 1 sources checked- cites [10] 2006
Not checked in the free run.
Not yet classified.
FindingThe responses of Cynomolgus macaque lymphocytes to immobilized TGN1412 plus IL-2 were only about 10% of human lymphocyte responses to immobilized TGN1412 alone or macaque lymphocytes stimulated with mitogen.
“However, the responses of Cynomolgus macaque lymphocytes to immobilized TGN1412 plus IL-2 were only ϳ10% of the responses of human lymphocytes to immobilized TGN1412 alone or of Cynomolgus macaque lymphocytes stimulated with mitogen”
Quote in the paperNot yet classifiedNot yet classified.
FindingTGN1412 air-dried onto wells stimulated release of TNF-alpha, IL-6, and IL-8 from human whole blood.
“When human whole blood at a final dilution of 20% (v/v) was substituted for PBMC, TGN1412 (1.0 and 10 g/well, but not 0.1 g/well) air dried onto the walls of wells in which 20% (v/v) human whole blood was then cultured for 24-h stimulated release of TNF-␣, IL-6, and IL-8”
Quote in the paperNot yet classifiedNot yet classified.
InterpretationThe activation of macaque lymphocytes by immobilized TGN1412 confirms binding to and interaction with macaque CD28.
“However, macaque lymphocytes were activated by immobilized TGN1412 (as shown by the up-regulation of IL-2R expression and blast transformation), confirming binding to and interaction with macaque CD28.”
Quote in the paperNot yet classifiedNot yet classified.
InterpretationThe 0.1 mg/kg dose given to volunteers in the clinical trial appears to have been close to the maximum immunostimulatory dose for lymphocyte proliferation.
“However, the dose of TGN1412 given to volunteers in the clinical trial (0.1 mg/kg) would appear to have been close to the maximum immunostimulatory dose for lymphocyte proliferation.”
Quote in the paperNot yet classified- Rests on an assumption: The in vitro concentration of TGN1412 can be converted to an equivalent in vivo dose in humans.
May combine two claims.
InterpretationreasoningTGN1412 is superagonistic in humans but not in Cynomolgus macaques.
“Essentially, TGN1412 is superagonistic in humans but not in Cynomolgus macaques.”
Quote in the paperSubstantial claimMay combine two claims.
InterpretationreasoningThe profound difference in responses of human and macaque lymphocytes to immobilized TGN1412 appears to take place at the level of cell signaling pathways, because TGN1412 binds avidly to and activates CD28+ lymphocytes from Cynomolgus macaques.
“Thus, the profound difference in responses of human and macaque lymphocytes to immobilized TGN1412 appears to take place at the level of cell signaling pathways, because TGN1412 binds avidly to and activates CD28 ϩ lymphocytes from Cynomolgus macaques.”
Quote in the paperSubstantial claimNot yet classified.
PremiseThe mouse anti-human CD28 mAb 5.11A1, from which TGN1412 was engineered, markedly stimulated human T cell proliferation in vitro with a bell-shaped dose-response curve.
“Consistent with our data, the superagonistic mouse anti-human CD28 mAb 5.11A1, from which TGN1412 was engineered, markedly stimulated human T cell proliferation in vitro, with a bell-shaped dose-response curve (11)”
Quote in the paperNot yet classified0 of 1 sources checked- cites [11] 2003
Not checked in the free run.
May combine two claims.
Study design and conductnone givenThe dose equivalence calculation relies on extrapolation from an in vivo dose to an in vitro concentration and from an in vivo human study to an in vitro study using isolated human white blood cells.
“Obviously, this calculation relies upon an extrapolation from an in vivo dose to an in vitro concentration and, more fundamentally, from an in vivo study in humans to an in vitro study using isolated human white blood cells.”
Quote in the paperSubstantial claimThe quote only partly states this claim.
Study design and conductown dataThe reported cytokine release results are representative of other independent experiments with different donors and conditions.
“The results of the above experiments are representative of the results of other independent experiments conducted with cells or”
- Also stated in: § Results · plain
“blood from other donors but using different cell densities, incubation times, or other changes in experimental conditions.”
Quote in the paperSubstantial claim- Also stated in: § Results · plain
May combine two claims.
PremisecitationHuman and Cynomolgus macaque CD28 extracellular domains are 100% identical, but this may not exclude functional differences.
“As previously reported, human and Cynomolgus macaque CD28 extracellular domains are 100% identical, but this may not exclude functional differences (12).”
Quote in the paperSubstantial claim0 of 1 sources checked- cites [12] 2006
Not checked in the free run.
May combine two claims.
Premisecitation§ "Cytokine Storm" in the Phase I Trial of Monoclonal Antibody TGN1412: Better Understanding the Causes to Improve PreClinical Testing of ImmunotherapeuticsAll six healthy volunteers in the Phase I trial of TGN1412 experienced life-threatening severe adverse events secondary to a cytokine storm and required weeks of hospitalization.
“The CD28-specific mAb TGN1412 rapidly caused a life-threatening”
- Also stated in: § A · plain
“All six recipients of the Ab suffered life-threatening severe adverse events, secondary to a”
Quote in the paperSubstantial claim0 of 1 sources checked- cites [1] 2006
Not checked in the free run.
- Also stated in: § A · plain
Not yet classified.
Study design and conductFor human proliferative responses, n=8; for macaque proliferative responses, n=8; the data were obtained in two independent experiments.
“For human proliferative responses, n ϭ 8.For macaque proliferative responses, n ϭ 8.The data were obtained in two independent experiments.”
Quote in the paperNot yet classifiedClassified as probably not a substantial claim. May combine two claims.
PremisecitationMacaque CD28 differs from human CD28 in three transmembrane residues, and it has been suggested that this could alter associations with molecular partners.
“Moreover, macaque CD28 differs from human CD28 in three transmembrane residues and it has been suggested that this could alter associations with molecular partners (13).”
Quote in the paperSubstantial claim0 of 1 sources checked- cites [13] 2006
Not checked in the free run.
Probably not a substantial claim: may not pass S3 (load-bearing).
Study design and conductown dataCytokine release data were obtained in three independent experiments with PBMC from different donors.
“The data were obtained in three independent experiments, each comprising PBMC from different donors: four, four, and three donors, respectively.”
Quote in the paperNot a substantial claim
The paper relies on this without support.
The maximum dose of TGN1412 given to macaques was 50 mg/kg, which is 500 times the human dose.
“TGN1412 was injected into volunteers at 0.1 mg/kg and into monkeys at up to the much higher dose of 50 mg/kg, i.e., 500ϫ the human dose.”
The paper relies on this without support.
The in vitro presentation of TGN1412 to PBMC that elicits cytokine release and proliferation mimics the in vivo presentation that caused the clinical toxicity.
“they did not present the Ab to white blood cells in a manner that mimicked its presentation in vivo.”
The paper relies on this without support.
The in vitro concentration of TGN1412 can be converted to an equivalent in vivo dose in humans.
“this calculation relies upon an extrapolation from an in vivo dose to an in vitro concentration and, more fundamentally, from an in vivo study in humans to an in vitro study using isolated human white blood cells.”
Claims other papers make about this one
Each is checked against this paper’s own text: does it say what the citing paper says it does?
Subsequent analysis of the TGN1412 used at the trial showed that it was formulated correctly, complied with its product specification and was free from pro-inflammatory contaminants.
“Subsequent analysis of the TGN1412 used at the trial showed that it was formulated correctly, complied with its product specification and was free from pro-inflammatory contaminants (Duff, 2006;Stebbings et al., 2007).”
From Monoclonal antibody TGN1412 trial failure explained by species differences in CD28 expression on CD4 <sup>+</sup> effector memory T‐cells (2010) · cites this paper as Stebbings et al., 2007
The introductory section (page not available) says TGN1412 used in the trial tested negative for endotoxin and describes subsequent analysis at NIBSC. The passages available from our copy do not establish that the trial material was correctly formulated, met its product specification, or was free of all pro-inflammatory contaminants; a negative endotoxin test supports only the narrower contamination claim. (LLM, conf=0.84)
Experimental — accuracy not yet measuredPBMCs from cynomolgus macaques were not stimulated in vitro by immobilized TGN1412, in agreement with the failure of in vivo administration at any dose tested to evoke any adverse response.
“Significantly, PBMCs from cynomolgus macaques were not stimulated in vitro by immobilized TGN1412 in agreement with the failure of in vivo administration at any dose tested to evoke any adverse response (Duff, 2006;Stebbings et al., 2007).”
From Monoclonal antibody TGN1412 trial failure explained by species differences in CD28 expression on CD4 <sup>+</sup> effector memory T‐cells (2010) · cites this paper as Stebbings et al., 2007
The Lymphocyte proliferation section reports very little proliferation of cynomolgus macaque lymphocytes with immobilized TGN1412, and the Discussion says all tested in vivo doses were well tolerated. But the Activation and intracellular cytokine staining section reports significant up-regulation of an activation marker in macaque T cells. Thus, saying the cells were not stimulated at all overstates the in vitro finding. Page numbers are unavailable in our copy. (LLM, conf=0.88)
Experimental — accuracy not yet measuredImmobilized TGN1412 primed macaque CD4+ T-cells, and they underwent marked lymphoproliferation only when exogenous IL-2 was added.
“However, we previously reported that immobilized TGN1412 primed macaque CD4 + T-cells and that they underwent marked lymphoproliferation only when exogenous IL-2 was added (Stebbings et al., 2007).”
From Monoclonal antibody TGN1412 trial failure explained by species differences in CD28 expression on CD4 <sup>+</sup> effector memory T‐cells (2010) · cites this paper as Stebbings et al., 2007
The Activation and intracellular cytokine staining section reports that immobilized TGN1412 activated macaque CD4+ T cells without inducing proliferation. The Lymphocyte proliferation section reports proliferation after recombinant IL-2 was added, but says that response was only about 10% of the human response or the macaque mitogen response; calling it “marked” overstates the finding. (LLM, conf=0.87)
Experimental — accuracy not yet measuredStandard in vitro safety tests with human cells were non-predictive because they did not replicate in vivo presentation of TGN1412.
“Standard in vitro safety tests with human cells were also non-predictive as they did not replicate in vivo presentation of TGN1412.”
From After TGN1412: Recent developments in cytokine release assays (2013) · cites this paper as Stebbings et al., 2007
The methods section reports that protocols similar to the original preclinical tests failed to elicit the cytokine release seen in volunteers, while immobilized TGN1412 did elicit responses in human cells. The Discussion proposes cell-surface binding as a requirement for the in vivo reaction, but labels this a hypothesis; it does not establish that failure to replicate the precise in vivo presentation caused the tests to be non-predictive. Our copy provides no page number for these passages. (LLM, conf=0.84)
Experimental — accuracy not yet measuredThe TGN1412 trial material was not contaminated and had been correctly formulated.
“Initially a contaminant was suspected of causing the adverse drug responses, but subsequent analysis showed that the trial material was not contaminated and had been correctly formulated (Duff, 2006; Stebbings et al., 2007).”
From After TGN1412: Recent developments in cytokine release assays (2013) · cites this paper as Stebbings et al., 2007
The introductory section (page unavailable) says the TGN1412 used in the trial tested negative for endotoxin contamination and came from a pharmaceutical-grade batch. It does not establish that the trial material was free of all contaminants or that it had been correctly formulated. (LLM, conf=0.86)
Experimental — accuracy not yet measured
The calculation of a safe starting dose in man based upon pre-clinical safety testing of TGN1412 in cynomolgus macaques was flawed.
“Moreover, the calculation of a safe starting dose in man based upon pre-clinical safety testing of TGN1412 in cynomolgus macaques was flawed.”
From After TGN1412: Recent developments in cytokine release assays (2013) · cites this paper as Stebbings et al., 2007
The Discussion says the cynomolgus macaque preclinical study failed to predict TGN1412 toxicity: doses tolerated by macaques included doses far above the 0.1 mg/kg dose toxic to human volunteers. This supports the conclusion that macaque safety testing did not establish a safe human starting dose, but the paper does not identify a flaw in the starting-dose calculation itself. No page number is available in our copy. (LLM, conf=0.82)
Experimental — accuracy not yet measuredThe 0.1 mg/kg dose correlated with 45-80% receptor occupancy in man and a near maximum immunostimulatory dose in vitro.
“In reality, this dose correlated with 45–80% receptor occupancy in man and a near maximum immunostimulatory dose in vitro (Stebbings et al., 2007; Waibler et al., 2008).”
From After TGN1412: Recent developments in cytokine release assays (2013) · cites this paper as Stebbings et al., 2007
The Discussion (page unavailable) says the 0.1 mg/kg human dose appeared close to the maximum immunostimulatory dose for lymphocyte proliferation in vitro, while noting that this rests on an in vivo-to-in vitro extrapolation. The passages on dosing and dose responses do not report 45–80% receptor occupancy in humans, so that numerical part is unsupported by this paper. (LLM, conf=0.88)
Experimental — accuracy not yet measuredTGN1412 stimulates CD4+ T-cells to undergo lymphoproliferation and release pro-inflammatory cytokines, whereas Campath-1H and Rituximab stimulate cytokine release from NK and CD8+, but not CD4+, T-cells.
“It was established that TGN1412 stimulates CD4+ T-cells to undergo lymphoproliferation and release pro-inflammatory cytokines (Stebbings et al., 2007).”
From After TGN1412: Recent developments in cytokine release assays (2013) · cites this paper as Stebbings et al., 2007
The Lymphocyte proliferation section identifies CD4+ T cells as the principal proliferating population, and Activation and intracellular cytokine staining reports IL-2 and IFN-γ production by human CD4+ lymphocytes. These findings were obtained with immobilized TGN1412 in vitro; the paper reports no significant proliferation with TGN1412 in aqueous solution, a condition the citing sentence does not distinguish. (LLM, conf=0.84)
Experimental — accuracy not yet measuredWhen standard pre-clinical safety tests were independently repeated using recovered clinical trial material, no pro-inflammatory cytokine release could be detected either in vitro with human white blood cells or when infused into cynomolgus macaques.
“When standard pre-clinical safety tests were independently repeated, using recovered clinical trial material, no pro-inflammatory cytokine release could be detected either in vitro with human white blood cells or when infused into cynomolgus macaques (Duff, 2006; Stebbings et al., 2007).”
From After TGN1412: Recent developments in cytokine release assays (2013) · cites this paper as Stebbings et al., 2007
Split into 2 atoms — 0 confirmed, 2 partially supported, 0 disputed, 0 misattributed, 0 unverifiable. Expand the claim to see per-atom verdicts.
Experimental — accuracy not yet measuredThe pro-inflammatory response to TGN1412 in the solid phase assay is human specific, as white blood cells from cynomolgus macaques were not stimulated by TGN1412 in a solid phase assay at any dose.
“The possibility that this response was an artefact was ruled out by the observation that white blood cells from cynomolgus macaques were not stimulated by TGN1412 in a solid phase assay at any dose 4 . The pro-inflammatory response to TGN1412 was human specific.”
From Severity of the <scp>TGN</scp>1412 trial disaster cytokine storm correlated with <scp>IL</scp>‐2 release (2013) · cites this paper as [4]
The Results (page unavailable) report that solid-phase TGN1412 induced pro-inflammatory cytokine release from human blood but not significant TNF-α, IL-6, or IL-8 release from cynomolgus macaque blood. However, the Lymphocyte proliferation and Discussion sections report some macaque-cell activation by immobilized TGN1412, so saying the cells were not stimulated at any dose overstates the finding. The species difference also does not, by itself, establish that an assay artefact was ruled out. (LLM, conf=0.86)
Experimental — accuracy not yet measuredIn the authors' in vitro assays, Mabthera and Campath-1H did not induce lymphoproliferation or IL-2 release, which are cardinal features of TGN1412 stimulation.
“When we investigated other therapeutic mAbs, known to be capable of causing adverse reactions, Mabthera and Campath-1H (Wing et al., 1995;Chung, 2008), using our new in vitro procedures, we did not detect lymphoproliferative responses or IL-2 release, the cardinal features of TGN1412 stimulation (Stebbings et al., 2007).”
From Monoclonal antibody TGN1412 trial failure explained by species differences in CD28 expression on CD4 <sup>+</sup> effector memory T‐cells (2010) · cites this paper as Stebbings et al., 2007
Following the TGN1412 incident, the authors developed new pre-clinical in vitro procedures using human PBMCs which can be applied to therapeutic mAbs during development, in order to prevent a repeat of the TGN1412 incident.
“Following the TGN1412 incident, we developed new pre-clinical in vitro procedures using human PBMCs which can be applied to therapeutic mAbs during development, in order to prevent a repeat of the TGN1412 incident (Stebbings et al., 2007;Findlay et al., 2010).”
From Monoclonal antibody TGN1412 trial failure explained by species differences in CD28 expression on CD4 <sup>+</sup> effector memory T‐cells (2010) · cites this paper as Stebbings et al., 2007
The in vitro safety procedures of Stebbings et al. (2007) are applicable to other therapeutic mAbs if IL-2 and IFN-g release are used as biomarkers for a TGN1412-type adverse response.
“Overall, these findings demonstrate that the in vitro safety procedures described by Stebbings et al. (2007) are applicable to other therapeutic mAbs, if IL-2 and IFN-g release are used as biomarkers for a TGN1412-type adverse response.”
From Monoclonal antibody TGN1412 trial failure explained by species differences in CD28 expression on CD4 <sup>+</sup> effector memory T‐cells (2010) · cites this paper as Stebbings et al. (2007)
Macaque T-cells do not secrete IL-2 when stimulated with TGN1412.
“Because macaque T-cells do not secrete IL-2 when stimulated with TGN1412 (Stebbings et al., 2007), it could be speculated that production of IL-2 by stimulated CD4 + effector memory T-cells is a missing factor required for in vitro lymphoproliferative responses by macaque CD4 + memory T-cells.”
From Monoclonal antibody TGN1412 trial failure explained by species differences in CD28 expression on CD4 <sup>+</sup> effector memory T‐cells (2010) · cites this paper as Stebbings et al., 2007
Central memory CD4+ T-cells are characterized by strong lymphoproliferative responses, which TGN1412 stimulates from human but not macaque PBMCs.
“Central memory CD4 + T-cells are characterized by strong lymphoproliferative responses (Sallusto et al., 1999;2004), which TGN1412 stimulates from human, but not macaque PBMCs (Stebbings et al., 2007).”
From Monoclonal antibody TGN1412 trial failure explained by species differences in CD28 expression on CD4 <sup>+</sup> effector memory T‐cells (2010) · cites this paper as Stebbings et al., 2007
If an immobilized therapeutic mAb-based assay or endothelial cell co-culture assay was used to evaluate TGN1412, then these would have predicted a pro-inflammatory response in man.
“It was subsequently shown that, if an immobilized therapeutic mAb-based assay or endothelial cell co-culture assay was used to evaluate TGN1412, then these would have predicted a pro-inflammatory response in man.”
From After TGN1412: Recent developments in cytokine release assays (2013) · cites this paper as Stebbings et al., 2007
Standard pre-clinical safety tests and animal models applied to TGN1412 had poor predictive value for clinical outcome in man.
“The poor predictive value of standard pre-clinical safety tests and animal models applied to TGN1412 demonstrated the need for a new generation of immunotoxicity assays and animal models that are both sensitive and predictive of clinical outcome in man.”
From After TGN1412: Recent developments in cytokine release assays (2013) · cites this paper as Stebbings et al., 2007
Immobilizing TGN1412 onto tissue culture plates so that human T-cells could only interact with the mAb at one interface resulted in massive lymphoproliferation and pro-inflammatory cytokine release that mirrored trial volunteer responses to TGN1412.
“The result of this simple change in presentation was dramatic; massive lymphoproliferation and pro-inflammatory cytokine release that mirrored trial volunteer responses to TGN1412 (Stebbings et al., 2007; Figure 2).”
From After TGN1412: Recent developments in cytokine release assays (2013) · cites this paper as Stebbings et al., 2007
Coating TGN1412 onto tissue culture wells in a solid phase assay caused massive pro-inflammatory cytokine release and lymphoproliferation that mirrored the responses to TGN1412 observed in trial volunteers.
“The result of this simple change in presentation was dramatic – massive pro-inflammatory cytokine release and lymphoproliferation that mirrored the responses to TGN1412 observed in trial volunteers 4 .”
From Severity of the <scp>TGN</scp>1412 trial disaster cytokine storm correlated with <scp>IL</scp>‐2 release (2013) · cites this paper as [4]
Pre-clinical safety tests repeated with the same trial material did not predict an adverse response in man.
“pre-clinical safety tests, repeated with the same trial material, did not predict an adverse response in man 3 – 4 .”
From Severity of the <scp>TGN</scp>1412 trial disaster cytokine storm correlated with <scp>IL</scp>‐2 release (2013) · cites this paper as [4]
The solid phase assay induces substantial cytokine release equivalent to that reported in trial volunteers, although it does not replicate in vivo presentation of TGN1412.
“Although the solid phase assay does not replicate in vivo presentation of TGN1412, which is not fully understood, it does induce substantial cytokine release equivalent to that reported in trial volunteers.”
From Severity of the <scp>TGN</scp>1412 trial disaster cytokine storm correlated with <scp>IL</scp>‐2 release (2013) · cites this paper as [4]
Experimental — accuracy not yet measuredA similar cytokine release profile was obtained when TGN1412 (0.1, 1.0, and 10 μg/well) was added in aqueous solution to a coculture of human PBMCs and endothelial cells.
“Also, a similar cytokine release profile was obtained when TGN1412 (0.1, 1.0, and 10 μ g/well) was added in aqueous solution to a coculture of human PBMCs and endothelial cells.”
From The calm after the cytokine storm: lessons from the TGN1412 trial (2008) · cites this paper as [11]
Stebbings et al. could not duplicate these responses using PBMCs from cynomolgus monkeys, suggesting that TGN1412 does not behave the same in nonhuman primate species as in humans.
“Interestingly, they could not duplicate these responses using PBMCs from cynomolgus monkeys, suggesting that TGN1412 does not behave the same in nonhuman primate species as in humans.”
From The calm after the cytokine storm: lessons from the TGN1412 trial (2008) · cites this paper as [11]
Stebbings et al. found that when TGN1412 (1.0 and 10.0 μg/well) was air dried on wells and added to human PBMCs, it induced the release of large amounts of TNFα, IL-6, and IL-8.
“They found that when TGN1412 (1.0 and 10.0 μ g/well) was air dried on wells and added to human PBMCs, it induced the release of large amounts of TNFα , IL-6, and IL-8.”
From The calm after the cytokine storm: lessons from the TGN1412 trial (2008) · cites this paper as [11]
TGN1412 must be artificially immobilized on the plastic surface of a culture well to induce cytokine release in vitro.
“In order for cytokine release to be induced, TGN1412 needs to be artificially immobilized on the plastic surface of the cell culture well [42]”
From The rise and fall of the <scp>CD</scp> 28 superagonist <scp>TGN</scp> 1412 and its return as <scp>TAB</scp> 08: a personal account (2016) · cites this paper as [42]
The plastic-immobilization protocol produced TGN1412 responses that mirrored the phase I cytokine release syndrome well, indicating that peripheral blood mononuclear cells contain cells capable of mounting an in vitro cytokine storm under artificial antibody presentation.
“the plastic-immobilization protocol championed by Stebbings and colleagues allowed TGN1412 responses which mirrored the CRS of the phase I study quite well [42], indicating that there are cells present in PBMC capable of responding to TGN1412 with an in vitro cytokine storm, albeit under artificial conditions of antibody presentation.”
From The rise and fall of the <scp>CD</scp> 28 superagonist <scp>TGN</scp> 1412 and its return as <scp>TAB</scp> 08: a personal account (2016) · cites this paper as [42]
Self-check
Percentages, sums and reported differences re-computed from the paper’s own numbers.
Each contradiction between two passages listed here was confirmed by an independent check.
Figure 7B gives conflicting units for the peak dose range
MinorThe results give the peak IL-2 response as 0.4-2 g/well, but the Figure 7B caption labels that same range as g/ml. The caption also gives 2-10 g/ml as the corresponding concentration range.
“Peak dose response for human lymphocytes was between 0.4 and 2 g/ml (2-10 g/ml).”
“For IL-2 production measured by intracellular cytokine staining, the optimal dose response of human lymphocytes to immobilized TGN1412 was between 0.4 and 2 g/well (Fig. 7B).”
IL-2 result points to the wrong figure
MinorThe cited Figure 5C-D concerns macaque activation, not the human IL-2 comparison described in the sentence. Figure 6C-D depicts the stated comparison.
“Even higher levels of IL-2 ϩ human lymphocytes were obtained with TGN142 captured in aqueous solution by immobilized anti-human Fc Ab compared with controls (Fig. 5, C and D)”
“Figure: 6 FIGURE 6 . FIGURE 6. Intracellular cytokine staining of human lymphocytes for IL-2 following a 6-h stimulation with (A) negative control, (B) 1 g/well immobilized TGN1412, (C) a combination of 1 g/well aqueous TGN1412 and 1 g/well aqueous anti-human Fc Ab, (D) 1 g/well aqueous TGN1412 captured by 1 g/well immobilized anti-human Fc Ab.”
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“Cytokine release assay To investigate the risk of systemic cytokine release in patients induced by PRS-343, an in vitro cytokine release assay (31) was carried out with PBMCs from 12 healthy control donors.”
MethodVafa, O., Gilliland, G. L., Brezski, R. J., Strake, B., Wilkinson, T., Lacy, E. R., Scallon, B. J., Teplyakov, A., Malia, T., & Strohl, W. R. (2013) · Methods · doi:10.1016/j.ymeth.2013.06.035 · Cited by 154 · see citing works
The Role of Fc Receptors on the Effectiveness of Therapeutic Monoclonal Antibodies
Gogesch, P., Dudek, S., van Zandbergen, G., Waibler, Z., & Anzaghe, M. (2021) · International Journal of Molecular Sciences · doi:10.3390/ijms22168947 · Cited by 152 · see citing works
Skokos, D., Waite, J. C., Haber, L., Crawford, A., Hermann, A., Ullman, E., Slim, R. M., Godin, S., Ajithdoss, D. K., Ye, X., et al. (2020) · Science Translational Medicine · doi:10.1126/scitranslmed.aaw7888 · Cited by 149 · see citing works
“S6) (30), we first used the FDA-recommended in vitro dry- and wet-coated human T cell proliferation assays (38) to determine whether either of our costimulatory bispecific antibodies could promote T cell activation and cytokine release in a manner equivalent to in-house generated CD28-SA (table S1).”
MethodResultInnovations, challenges, and minimal information for standardization of humanized mice
Stripecke, R., Münz, C., Schuringa, J. J., Bissig, K., Soper, B. W., Meeham, T., Yao, L., Santo, J. P. D., Brehm, M. A., Rodríguez, E., et al. (2020) · EMBO Molecular Medicine · doi:10.15252/emmm.201708662 · Cited by 136 · see citing works
“Dendritic cells (DCs) are main orchestrators of the adaptive immune system presenting processed peptide antigens to T cells through MHC classes I/II and expressing key costimulatory molecules such as CD40 ligand (CD40L) required for B-cell activation and class switching (Steinman, 2012).”
BackgroundClinical development methodology for infusion‐related reactions with monoclonal antibodies
Doessegger, L., & Banholzer, M. L. (2015) · Clinical & Translational Immunology · doi:10.1038/cti.2015.14 · Cited by 127 · see citing works
Golay, J., & Introna, M. (2012) · Archives of Biochemistry and Biophysics · doi:10.1016/j.abb.2012.02.011 · Cited by 127 · see citing works
Translating costimulation blockade to the clinic: lessons learned from three pathways
Ford, M. L., & Larsen, C. P. (2009) · Immunological Reviews · doi:10.1111/j.1600-065x.2009.00776.x · Cited by 123 · see citing works
“hyperactivated and secrete inflammatory cytokines (27).”
Burova, E., Hermann, A., Waite, J. C., Potocky, T. B., Lai, V. K.-M., Hong, S., Liu, M., Allbritton, O., Woodruff, A., Wu, Q., et al. (2017) · Molecular Cancer Therapeutics · doi:10.1158/1535-7163.mct-16-0665 · Cited by 115 · see citing works
“To determine whether REGN2810 by itself stimulates T cells in the absence of TCR activation, we employed a cytokine release assay (45).”
MethodCytokine release syndrome and cancer immunotherapies – historical challenges and promising futures
Shah, D., Soper, B. W., & Shopland, L. S. (2023) · Frontiers in Immunology · doi:10.3389/fimmu.2023.1190379 · Cited by 116 · see citing works
Immune-Mediated Adverse Drug Reactions
Uetrecht, J. (2008) · Chemical Research in Toxicology · doi:10.1021/tx800389u · Cited by 109 · see citing works
Non-immunotherapy application of LNP-mRNA: Maximizing efficacy and safety
Vlatkovic, I. (2021) · Biomedicines · doi:10.3390/biomedicines9050530 · Cited by 112 · see citing works
Safety and secondary pharmacology: Successes, threats, challenges and opportunities
Valentin, J., & Hammond, T. G. (2008) · Journal of Pharmacological and Toxicological Methods · doi:10.1016/j.vascn.2008.05.007 · Cited by 110 · see citing works
“Nevertheless, recent investigations on the mechanisms of the TGN-1412mediated “cytokine storm” will enable to develop novel procedures to improve non-clinical safety testing of immunomodulatory therapeutics (Stebbings et al., 2007).”
BackgroundCytokine release assays: Current practices and future directions
Finco, D., Grimaldi, C., Fort, M., Walker, M., Kiessling, A., Wolf, B., Salcedo, T., Faggioni, R., Schneider, A., Ibraghimov, A., et al. (2014) · Cytokine · doi:10.1016/j.cyto.2013.12.009 · Cited by 106 · see citing works
Self-citationHynds, R. E., & Giangreco, A. (2012) · Stem Cells · doi:10.1002/stem.1290 · Cited by 105 · see citing works
Muller, P., & Brennan, F. R. (2009) · Clinical Pharmacology & Therapeutics · doi:10.1038/clpt.2008.273 · Cited by 104 · see citing works
Ramani, T., Auletta, C. S., Weinstock, D., Mounho-Zamora, B., Ryan, P. C., Salcedo, T. W., & Bannish, G. (2015) · International Journal of Toxicology · doi:10.1177/1091581815584918 · Cited by 103 · see citing works
Tabares, P., Berr, S., Römer, P. S., Chuvpilo, S., Matskevich, A. A., Tyrsin, D., Fedotov, Y., Einsele, H., Tony, H., & Hünig, T. (2014) · European Journal of Immunology · doi:10.1002/eji.201343967 · Cited by 97 · see citing works
Immunological substance testing on human lymphatic micro-organoids in vitro
Giese, C., Lubitz, A., Demmler, C. D., Reuschel, J., Bergner, K., & Marx, U. (2010) · Journal of Biotechnology · doi:10.1016/j.jbiotec.2010.03.001 · Cited by 95 · see citing works
Differential Monocyte Actuation in a Three‐Organ Functional Innate Immune System‐on‐a‐Chip
Sasserath, T., Rumsey, J. W., McAleer, C. W., Bridges, L. R., Long, C. J., Elbrecht, D. H., Schuler, F., Roth, A. B., Bertinetti‐LaPatki, C., Shuler, M. L., et al. (2020) · Advanced Science · doi:10.1002/advs.202000323 · Cited by 93 · see citing works
Horvath, C. J., & Milton, M. N. (2009) · Toxicologic Pathology · doi:10.1177/0192623309332986 · Cited by 87 · see citing works
“Within ninety minutes, those who had received TGN1412 had a systemic inflammatory response that was characterized by a rapid induction of proinflammatory cytokines and accompanied by headache, myalgia, nausea, diarrhea, erythema, vasodilatation, and hypotension (Suntharalingam et al. 2006).”
BackgroundResultInfluentialRao, D. A., Eid, R. E., Qin, L., Yi, T., Kirkiles-Smith, N. C., Tellides, G., & Pober, J. S. (2008) · The Journal of Experimental Medicine · doi:10.1084/jem.20081661 · Cited by 86 · see citing works
Agoram, B. M. (2008) · British Journal of Clinical Pharmacology · doi:10.1111/j.1365-2125.2008.03297.x · Cited by 85 · see citing works
Harper, J. V., Adams, K. J., Bossi, G., Wright, D. E., Stacey, A., Bedke, N., Martinez-Hague, R., Blat, D., Humbert, L., Buchanan, H., et al. (2018) · PLoS ONE · doi:10.1371/journal.pone.0205491 · Cited by 84 · see citing works
Unraveling the Effect of Immunogenicity on the PK/PD, Efficacy, and Safety of Therapeutic Proteins
Smith, A. M., Manoli, H., Jaw, S., Frutoz, K., Epstein, A. L., Khawli, L. A., & Theil, F. (2016) · Journal of Immunology Research · doi:10.1155/2016/2342187 · Cited by 80 · see citing works
“Immunogenicity-related adverse events (AEs) may be as benign as a mild itchy rash or as serious as infusion reactions, anaphylaxis, and cytokine release syndrome (CRS) [13, 14].”
Signaling Signatures and Functional Properties of Anti-Human CD28 Superagonistic Antibodies
Waibler, Z., Sender, L. Y., Merten, C., Hartig, R., Kliche, S., Gunzer, M., Reichardt, P., Kalinke, U., & Schraven, B. (2008) · PLoS ONE · doi:10.1371/journal.pone.0001708 · Cited by 79 · see citing works
“In line with previous data obtained using TGN1412 [32] Fig.”
ResultThe calm after the cytokine storm: lessons from the TGN1412 trial
St. Clair, E. W. (2008) · Journal of Clinical Investigation · doi:10.1172/jci35382 · Cited by 79 · see citing works
Poirier, N., Mary, C., Dilek, N., Hervouet, J., Minault, D., Blancho, G., & Vanhove, B. (2012) · American Journal of Transplantation · doi:10.1111/j.1600-6143.2012.04164.x · Cited by 79 · see citing works
“We therefore conducted a series of experiments following the recommendations published by the National Institute for Biological Standards and Control from the UK (58,59) after the TeGenero affair.”
BackgroundMethodFindlay, L., Eastwood, D., Stebbings, R., Sharp, G., Mistry, Y., Ball, C., Hood, J., Thorpe, R., & Poole, S. (2010) · Journal of Immunological Methods · doi:10.1016/j.jim.2009.10.013 · Cited by 74 · see citing works
Self-citationAfter TGN1412: Recent developments in cytokine release assays
Stebbings, R., Eastwood, D., Poole, S., & Thorpe, R. (2013) · Journal of Immunotoxicology · doi:10.3109/1547691x.2012.711783 · Cited by 73 · see citing works
“Predictivity of newly-proposed cytokine release assays Initial in vitro studies of pro-inflammatory cytokine release from human lymphocytes stimulated with immobilized TGN1412 measured high levels of tumor necrosis factor (TNF)-α and interleukin (IL)-8 (Stebbings et al., 2007).”
BackgroundMethodInfluentialSelf-citationConsiderations for the development of therapeutic monoclonal antibodies
Swann, P. G., Tolnay, M., Muthukkumar, S., Shapiro, M. A., Rellahan, B. L., & Clouse, K. A. (2008) · Current Opinion in Immunology · doi:10.1016/j.coi.2008.05.013 · Cited by 73 · see citing works
CD28 co-stimulation: novel insights and applications in cancer immunotherapy
Lotze, M. T., Olejniczak, S. H., & Skokos, D. (2024) · Nature reviews. Immunology · doi:10.1038/s41577-024-01061-1 · Cited by 77 · see citing works
“These and other conclusions of the Expert Scientific Group formed the basis for revision of the guidelines for first-in-human trials with investigational medicinal products by the European Medicines Agency 201 .”
Veitonmäki, N., Hansson, M., Zhan, F., Sundberg, A., Löfstedt, T., Ljungars, A., Li, Z., Niskanen, T. M., Zeng, M., Yang, Y., et al. (2013) · Cancer Cell · doi:10.1016/j.ccr.2013.02.026 · Cited by 72 · see citing works
“In order tomaximize the chances of identifying any PBMCagonistic properties of BI-505, we used two different antibodycoating protocols in which the antibody was hypercrosslinked as previously described (Stebbings et al., 2007).”
MethodSafety of biologics, lessons learnt from TGN1412
Stebbings, R., Poole, S., & Thorpe, R. (2009) · Current Opinion in Biotechnology · doi:10.1016/j.copbio.2009.10.002 · Cited by 70 · see citing works
Self-citationPoirier, N., Blancho, G., Hiance, M., Mary, C., Assche, T. V., Leempoels, J., Ramael, S., Wang, W., Thépenier, V., Braudeau, C., et al. (2016) · The Journal of Immunology · doi:10.4049/jimmunol.1601538 · Cited by 66 · see citing works
“Before the initiation of the current phase I study in healthy volunteers, the antagonistic properties of FR104 had been assessed thoroughly in vitro, as previously recommended (33, 34) as well as in vivo using humanized mice (20) and baboons (27).”
BackgroundVessillier, S., Eastwood, D., Fox, B. J., Sathish, J. G., Sethu, S., Dougall, T., Thorpe, S. J., Thorpe, R., & Stebbings, R. (2015) · Journal of Immunological Methods · doi:10.1016/j.jim.2015.04.020 · Cited by 68 · see citing works
Self-citationEarly Implementation of QbD in Biopharmaceutical Development: A Practical Example
Zurdo, J., Arnell, A., Obrezanova, O., Smith, N., de la Cuesta, R. G., Gallagher, T. R. A., Michael, R., Stallwood, Y., Ekblad, C., Abrahmsén, L., et al. (2015) · BioMed Research International · doi:10.1155/2015/605427 · Cited by 66 · see citing works
“any sample contamination or errors in the manufacturing, formulation, dilution or administration of TGN1412 that led to the CRS response [63, 82–86].”
BackgroundPolson, A. G., & Fuji, R. N. (2012) · British Journal of Pharmacology · doi:10.1111/j.1476-5381.2012.01916.x · Cited by 66 · see citing works
“This acute cytokine reaction did not occur in the non-human primate study (Suntharalingam et al., 2006; Stebbings et al., 2007).”
BackgroundMonoclonal Antibody First Dose Cytokine Release Syndromes–Mechanisms and Prediction
Wing, M. G. (2008) · Journal of Immunotoxicology · doi:10.1080/15476910801897433 · Cited by 64 · see citing works
“However unlike the clinical situation where massive cytokine release was detected, cytokines could not be detected in vitro unless the antibody was immobilised by “dry coating” the plates with the product (Stebbings et al., 2007).”
BackgroundSurrogate end points in the design of immunotherapy trials: emerging lessons from type 1 diabetes
Roep, B. O., & Peakman, M. (2010) · Nature reviews. Immunology · doi:10.1038/nri2705 · Cited by 63 · see citing works
Ye, C., Yang, H., Cheng, M., Shultz, L. D., Greiner, D. L., Brehm, M. A., & Keck, J. (2020) · The FASEB Journal · doi:10.1096/fj.202001203r · Cited by 60 · see citing works
Human immunity in vitro — Solving immunogenicity and more
Giese, C., & Marx, U. (2014) · Advanced Drug Delivery Reviews · doi:10.1016/j.addr.2013.12.011 · Cited by 62 · see citing works
Literature review on in vitro and alternative Developmental Neurotoxicity (DNT) testing methods
Fritsche, E., Alm, H., Baumann, J., Geerts, L., Håkansson, H., Masjosthusmann, S., & Witters, H. E. (2015) · EFSA Supporting Publications · doi:10.2903/sp.efsa.2015.en-778 · Cited by 61 · see citing works
Jiménez, S. A., & Piera-Velázquez, S. (2013) · Autoimmunity Reviews · doi:10.1016/j.autrev.2013.04.004 · Cited by 60 · see citing works
Therapeutic T-cell manipulation in rheumatoid arthritis: past, present and future
Isaacs, J. D. (2008) · Lara D. Veeken · doi:10.1093/rheumatology/ken163 · Cited by 60 · see citing works
“The life-threatening cytokine storm experienced by the human volunteers was difficult to replicate in vitro even after the event and has resulted in further safeguards during mAb development [62, 63 ].”
Development of CDX-1140, an agonist CD40 antibody for cancer immunotherapy
Vitale, L. A., Thomas, L. J., He, L.-Z., et al. (2018) · Cancer Immunology Immunotherapy · doi:10.1007/s00262-018-2267-0 · Cited by 59 · see citing works
Gaspar, M., Pravin, J., Rodrigues, L., Uhlenbroich, S., Everett, K. L., Wollerton, F., Morrow, M., Tuna, M., & Brewis, N. (2020) · Cancer Immunology Research · doi:10.1158/2326-6066.cir-19-0798 · Cited by 58 · see citing works
“© 2020 American Association for Cancer Research. cancerimmunolres.aacrjournals.org Downloaded from control antibodies used, a CD28 antibody shown to induce cytokine storm in the clinic (TGN1412; ref.”
MethodTherapeutic Monoclonal Antibodies
Yamada, T. (2011) · The Keio Journal of Medicine · doi:10.2302/kjm.60.37 · Cited by 57 · see citing works
CD28 Superagonists: What Makes the Difference in Humans?
Schraven, B. L., & Kalinke, U. (2008) · Immunity · doi:10.1016/j.immuni.2008.04.003 · Cited by 58 · see citing works
Eastwood, D., Bird, C., Dilger, P., Hockley, J., Findlay, L., Poole, S., Thorpe, S. J., Wadhwa, M., Thorpe, R., & Stebbings, R. (2013) · British Journal of Clinical Pharmacology · doi:10.1111/bcp.12165 · Cited by 58 · see citing works
Self-citationUpregulation of FcγRIIb on monocytes is necessary to promote the superagonist activity of TGN1412
Hussain, K., Hargreaves, C. E., Roghanian, A., Oldham, R. J., Chan, H. T. C., Mockridge, C. I., Chowdhury, F., Frendéus, B., Harper, K., Strefford, J. C., et al. (2014) · Blood · doi:10.1182/blood-2014-08-593061 · Cited by 55 · see citing works
“The immobilization of TGN1412 onto plastic or the addition of antiimmunoglobulin G (IgG) antibody has been shown to induce cytokine release in PBMC assays.(6,7) Both approaches provide extensive cross-linking of the mAb on the T-cell surface.”
BackgroundMethodA Monovalent Anti-Human CD28 Domain Antibody Antagonist: Preclinical Efficacy and Safety
Suchard, S. J., Davis, P. M., Kansal, S., Stetsko, D. K., Brosius, R., Tamura, J. K., Schneeweis, L. A., Bryson, J. W., Salcedo, T. W., Wang, H., et al. (2013) · The Journal of Immunology · doi:10.4049/jimmunol.1300470 · Cited by 55 · see citing works
“(29) to demonstrate agonist activity of TGN1412 in vitro.”
BackgroundSchabowsky, R.-H., Elpek, K. G., Madireddi, S., Sharma, R. K., Yolcu, E. S., Bandura‐Morgan, L., Miller, R. E., MacLeod, K. J., Mittler, R. S., & Shirwan, H. (2009) · Vaccine · doi:10.1016/j.vaccine.2009.09.127 · Cited by 54 · see citing works
In vivoeffects of targeting CD79b with antibodies and antibody-drug conjugates
Zheng, B., Fuji, R. N., Elkins, K., et al. (2009) · Molecular Cancer Therapeutics · doi:10.1158/1535-7163.mct-09-0369 · Cited by 54 · see citing works
“In some cases, cynomolgus monkeys are not predictive of cytokine release in humans; however, this appears to be a property of the target and not a general property of cynomolgus monkeys (23).”
BackgroundBegum, J., Day, W. P., Henderson, C. E., et al. (2013) · Cytometry Part A · doi:10.1002/cyto.a.22403 · Cited by 52 · see citing works
“For example, we are also looking at the performance of lipophilic dyes in cell lines (36) using our metrics as although they are used successfully to track division (6,10), peak resolution is inferior compared with”
MethodFilby, A., Begum, J., Jalal, M., & Day, W. (2015) · Methods · doi:10.1016/j.ymeth.2015.02.016 · Cited by 53 · see citing works
An FDA oncology analysis of immune activating products and first-in-human dose selection
Saber, H., Gudi, R., Manning, M. L., Wearne, E., & Leighton, J. K. (2016) · Regulatory Toxicology and Pharmacology · doi:10.1016/j.yrtph.2016.10.002 · Cited by 53 · see citing works
In vitro models for immunogenicity prediction of therapeutic proteins
Groell, F., Jordan, O., & Borchard, G. (2018) · European Journal of Pharmaceutics and Biopharmaceutics · doi:10.1016/j.ejpb.2018.06.008 · Cited by 53 · see citing works
Grimaldi, C. M., Finco, D., Fort, M. M., Gliddon, D., Harper, K., Helms, W. S., Mitchell, J. A., O’Lone, R. B., Parish, S. T., Piché, M.-S., et al. (2016) · Cytokine · doi:10.1016/j.cyto.2016.06.006 · Cited by 52 · see citing works
Self-citationBailey, J., & Balls, M. (2019) · BMC Medical Ethics · doi:10.1186/s12910-019-0352-3 · Cited by 51 · see citing works
The laboratory’s role in combating COVID-19
Fang, B., & Meng, Q. H. (2020) · Critical Reviews in Clinical Laboratory Sciences · doi:10.1080/10408363.2020.1776675 · Cited by 48 · see citing works
Wolf, B., Morgan, H., Krieg, J., Gani, Z., Milicov, A., Warncke, M., Brennan, F. R., Jones, S., Sims, J., & Kießling, A. (2012) · Cytokine · doi:10.1016/j.cyto.2012.08.018 · Cited by 50 · see citing works
Landrigan, A., Wong, M. T., & Utz, P. J. (2011) · The Journal of Immunology · doi:10.4049/jimmunol.1003414 · Cited by 47 · see citing works
“Additionally, costimulation with ODNs is an attractive alternative to CD28 costimulation with respect to patient therapy, as anti-CD28 Abs have demonstrated life-threatening clinical toxicity (66).”
BackgroundHünig, T. (2016) · The FEBS Journal · doi:10.1111/febs.13754 · Cited by 47 · see citing works
Immunotherapeutic strategies in autoimmune uveitis
Papotto, P. H., Marengo, E. B., Sardinha, L. R., Goldberg, A. C., & Rizzo, L. V. (2014) · Autoimmunity Reviews · doi:10.1016/j.autrev.2014.05.003 · Cited by 46 · see citing works
A CD28 superagonistic antibody elicits 2 functionally distinct waves of T cell activation in rats
Müller, N., van den Brandt, J., Odoardi, F., Tischner, D., Herath, J., Flügel, A., & Reichardt, H. M. (2008) · Journal of Clinical Investigation · doi:10.1172/jci32698 · Cited by 46 · see citing works
Shi, R., Honczarenko, M., Zhang, S. X., et al. (2016) · The Journal of Clinical Pharmacology · doi:10.1002/jcph.791 · Cited by 45 · see citing works
Summary of a workshop on preclinical and translational safety assessment of CD3 bispecifics
Kamperschroer, C., Shenton, J. M., Lebrec, H. N., Leighton, J. K., Moore, P. A., & Thomas, O. (2020) · Journal of Immunotoxicology · doi:10.1080/1547691x.2020.1729902 · Cited by 45 · see citing works
Mechanisms of Adverse Drug Reactions to Biologics
Clarke, J. B. (2009) · Handbook of experimental pharmacology · doi:10.1007/978-3-642-00663-0_16 · Cited by 45 · see citing works
“However, the manner in which the TGN1412 antibody was presented to the PBMCs proved crucial because when the antibody was first air dried onto the plate, wet coated on top of anti-Fc antibodies, or coated on top of endothelial cells, all resulted in the release of cytokines into the medium and lymphocyte proliferation…”
BackgroundMethodDhir, V., Fort, M. M., Mahmood, A., Higbee, R. G., Warren, W., Narayanan, P. K., & Wittman, V. (2011) · Journal of Immunotoxicology · doi:10.3109/1547691x.2011.613419 · Cited by 44 · see citing works
“Interestingly, in the absence of endothelial cells and collagen matrix, “RL-100” culture, the SI and/or magnitude of response to TGN1412 was greatly reduced for all the cytokines (Table 1, Figure 1); this latter finding confirmed a need for F c R-mediate cross-linking to occur for efficient induction of proinflammator…”
BackgroundMethodResultInfluentialVidal, J.-M., Kawabata, T. T., Thorpe, R., Lima, B. S., Cederbrant, K., Poole, S., Mueller-Berghaus, J., Pallardy, M. J., & van der Laan, J. W. (2010) · Cytokine · doi:10.1016/j.cyto.2010.04.008 · Cited by 43 · see citing works
Self-citationBreaking through the therapeutic ceiling of inflammatory bowel disease: Dual-targeted therapies
Feng, Z., Kang, G., Wang, J., Gao, X., Wang, X., Ye, Y., Liu, L., Zhao, J., Liu, X., Huang, H., et al. (2022) · Biomedicine & Pharmacotherapy · doi:10.1016/j.biopha.2022.114174 · Cited by 46 · see citing works
TGN1412 Induces Lymphopenia and Human Cytokine Release in a Humanized Mouse Model
Weißmüller, S., Kronhart, S., Kreuz, D., Schnierle, B. S., Kalinke, U., Kirberg, J., Hanschmann, K.-M. O., & Waibler, Z. (2016) · PLoS ONE · doi:10.1371/journal.pone.0149093 · Cited by 43 · see citing works
Bartholomaeus, P., Semmler, L. Y., Bukur, T., Boisguerin, V., Römer, P. S., Tabares, P., Chuvpilo, S., Tyrsin, D. Y., Matskevich, A., Hengel, H., et al. (2014) · The Journal of Immunology · doi:10.4049/jimmunol.1302461 · Cited by 40 · see citing works
“T cells enriched from freshly isolated PBMCs are not strongly activated upon treatment with soluble superagonistic anti-CD28 mAb TGN1412 (8, 16).”
BackgroundApplication of microphysiological systems in biopharmaceutical research and development
Peterson, N. C., Mahalingaiah, P. K. S., Fullerton, A. M., & Piazza, M. (2020) · Lab on a Chip · doi:10.1039/c9lc00962k · Cited by 40 · see citing works
Primate Models in Organ TransplantationPreprint
Anderson, D. J., & Kirk, A. D. (2013) · Cold Spring Harbor Perspectives in Medicine · doi:10.1101/cshperspect.a015503 · Cited by 39 · see citing works
“Although several recommendations have been made for the design of future trials to avoid a repeat of the TGN141 trial (Stebbings et al. 2007; Findlay et al. 2010), there has been, understandably, extreme caution in developing antibodies against CD28.”
BackgroundStorm forecasting: additional lessons from the CD28 superagonist TGN1412 trial
Horvath, C. J., Andrews, L., Baumann, A., Black, L. E., Blanset, D. L., Cavagnaro, J. A., Hastings, K. L., Hutto, D., MacLachlan, T. K., Milton, M., et al. (2012) · Nature reviews. Immunology · doi:10.1038/nri3192-c1 · Cited by 37 · see citing works
Hedgecoe, A. M. (2013) · Social Studies of Science · doi:10.1177/0306312713506141 · Cited by 37 · see citing works
“In terms of biology, what went wrong swiftly became clear (Stebbings et al., 2007).”
BackgroundREPRINT: Inflammatory findings on species extrapolations: humans are definitely no 70-kg mice 1
Leist, M. (2013) · ALTEX · doi:10.14573/altex.2013.2.227 · Cited by 35 · see citing works
- Authors
- Stebbings, R., Findlay, L., Edwards, C., Eastwood, D., Bird, C., North, D., Mistry, Y., Dilger, P., Liefooghe, E., Cludts, I., Fox, B., Tarrant, G., Robinson, J., Meager, T., Dolman, C., Thorpe, S. J., Bristow, A., Wadhwa, M., Thorpe, R., & Poole, S.
- Affiliations
- National Institute for Biological Standards and Control (GB)
- Venue
- The Journal of Immunology
- Publisher
- Oxford University Press (OUP)
- Published
- 2007-09-01
- Volume
- 179
- Issue
- 5
- Pages
- 3325-3331
- ISSN
- 0022-1767 (ISSN-L)
- 1550-6606
- OpenAlex
- W1705666194
- Semantic Scholar
- eacb1c28d1345a05916a17b7d8cf11667ce5c89c
- Cited by
- 317
- References
- 13
- Citations by year
- 2026: 8 · 2025: 9 · 2024: 16 · 2023: 12 · 2022: 7 · 2021: 7 · 2020: 17 · 2019: 14 · 2018: 14 · 2017: 14 · 2016: 17 · 2015: 18 · 2014: 24 · 2013: 24 · 2012: 31
- FWCI
- 7.57
- Type
- journal article
- Language
- en
- Open access
- bronze
- doi.org
- Locations
- The Journal of Immunology (publishedVersion)
- PubMed (publishedVersion)
- Topics
- Life Sciences › Immunology and Microbiology › Immunology › T-cell and B-cell Immunology
- Life Sciences › Immunology and Microbiology › Immunology › Immunotherapy and Immune Responses
- Life Sciences › Immunology and Microbiology › Immunology › Immune Cell Function and Interaction
- Keywords
- TGN1412, cytokine release syndrome, superagonists, preclinical safety, immunotherapeutics, lymphocyte proliferation, in vitro toxicity testing, non-human primates
- MeSH terms
- Animals, Antibodies, Monoclonal, Drug Evaluation, Preclinical, Humans, Immunotherapy, Leukocytes, Mononuclear
About these checks
Registry, retraction, Cites and statistics results are computed, not judged, with one exception: a printed reference that no index lists is read once by a model before it counts as a citation error. Claims are extracted by a model and kept only when their quote appears verbatim in the text; each cited source is then checked, with a model’s help, for whether it says what the claim relies on. Those verdicts can be wrong — read the claim’s quote and the cited source before relying on one.
Claims read by deepseek-v4.1-flash under definition version 3 (extract-v3) on 2026-09-28.
Classified by jev-1.13-20260917 (gold-guideline-v0.2) on 2026-09-28.
Statistics and arithmetic are recomputed, not judged.
Confidence bands are shown only for fields calibrated on our annotated test set. None is calibrated yet, so no band is shown.
What counts as a claim ›How we measure confidence ›How we check this ›
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