Development of an IFN-&amp;#947; ELISpot Assay to Assess Varicella-Zoster Virus-specific Cell-mediated Immunity Following Umbilical Cord Blood Transplantation

Fourati, Insaf Salem; Grenier, Anne-Julie; Jolette, Élyse; Mérindol, Natacha; Ovetchkine, Philippe; Soudeyns, Hugo

doi:10.3791/51643

Cited by 6 publications

(5 citation statements)

References 44 publications

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“…It has been also reported that percentage of VZV‐specific CD8 + effector memory T cells is significantly lower in KT recipients than in controls . However, current methods for the enumeration of VZV‐specific T cells—based on in vitro release of interferon‐γ or other Th 1 cytokines upon antigen stimulation—are technically demanding and not standardized, and their implementation in clinical practice seems unfeasible in the medium term . Although limited by the lack of an independent validation cohort, our study suggests that the NK cell count may be used as a convenient surrogate for the VZV‐specific cell‐mediated immunity.…”

Section: Discussionmentioning

confidence: 79%

Herpes zoster in kidney transplant recipients: protective effect of anti-cytomegalovirus prophylaxis and natural killer cell count. A single-center cohort study

et al. 2017

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Despite its impact on quality of life and potential for complications, specific risk and protective factors for herpes zoster (HZ) after kidney transplantation (KT) remain to be clarified. We included 444 patients undergoing KT between November 2008 and March 2013. Peripheral blood lymphocyte subpopulations were measured at baseline and months 1 and 6. The risk factors for early (first post-transplant year) and late HZ (years 1-5) were separately assessed. We observed 35 episodes of post-transplant HZ after a median follow-up of 48.3 months (incidence rate: 0.057 per 1000 transplant-days). Median interval from transplantation was 18.3 months. Six patients (17.1%) developed disseminated infection. Postherpetic neuralgia occurred in 10 cases (28.6%). The receipt of anti-cytomegalovirus (CMV) prophylaxis with (val)ganciclovir decreased the risk of early HZ [adjusted hazard ratio (aHR): 0.08; 95% CI: 0.01-1.13; P-value = 0.062], whereas the natural killer (NK) cell at month 6 was protective for the occurrence of late HZ [aHR (per 10-cells/μl increase): 0.94; 95% CI: 0.88-1.00; P-value = 0.054]. In conclusion, two easily ascertainable factors (whether the patient is receiving anti-CMV prophylaxis and the NK cell count at month 6) might be potentially useful to tailor preventive strategies according to individual susceptibility to post-transplant HZ.

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Section: Discussionmentioning

confidence: 79%

Herpes zoster in kidney transplant recipients: protective effect of anti-cytomegalovirus prophylaxis and natural killer cell count. A single-center cohort study

et al. 2017

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“…The correlation between VZV-specific IFN-γ release and T-cell subsets suggests that IFN-γ is mainly secreted by T-cells. To gain the detection sensitivity of VZV-responsive T-cells at the single-cell level, the IFN-γ enzyme-linked immunosorbent spot (ELISPOT) assay has been proposed ( 9 , 31 , 32 , 38 ). However, in addition to being labor intensive because of the multiple experimental steps from PBMC preparation to spot detection, enumeration of IFN-γ spot-forming cells proves to be difficult.…”

Section: Discussionmentioning

confidence: 99%

“…HI (n = 17) and allo-HSCT recipients (n = 60) were included. HI and allo-HSCT recipients did not significantly differ in terms of age (median [IQR], 42 [33][34][35][36][37][38][39][40][41][42][43][44][45][46][47][48][49][50][51] vs. 44.5 years, P = .39) and sex (sex ratio, 1.1 vs. 1.3, P = .79). Allo-HSCT recipients were enrolled at a median [IQR] time of 6 [5][6][7][8] months posttransplant.…”

Section: Participant Characteristicsmentioning

confidence: 99%

A Simple-to-Perform ifn-γ mRNA Gene Expression Assay on Whole Blood Accurately Appraises Varicella Zoster Virus-Specific Cell-Mediated Immunity After Allogeneic Hematopoietic Stem Cell Transplantation

et al. 2022

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Herpes zoster, which is due to the reactivation of Varicella zoster virus (VZV), is a leading cause of morbidity after allogeneic hematopoietic stem cell transplantation (allo-HSCT). While cell-mediated immunity (CMI) is critical to inhibiting VZV reactivation, CMI is not routinely assessed due to a lack of reliable tests. In this study, we aimed to evaluate VZV-specific CMI among allo-HSCT recipients (n = 60) and healthy individuals (HI, n = 17) through a panel of three immune functional assays after ex vivo stimulation by VZV antigen: quantification of (i) IFN-γ release in the supernatants, (ii) T-cell proliferation after a 7-day stimulation of peripheral blood mononuclear cells (PBMC), and (iii) measurement of the ifn-γ mRNA gene expression level after 24 h of stimulation of a whole-blood sample. VZV responsiveness was defined according to IFN-γ release from VZV-stimulated PBMC. Upon VZV stimulation, we found that allo-HSCT recipients at a median time of 6 [5-8] months post-transplant had lower IFN-γ release (median [IQR], 0.34 [0.12–8.56] vs. 409.5 [143.9–910.2] pg/ml, P <.0001) and fewer proliferating T cells (0.05 [0.01–0.57] % vs. 8.74 [3.12–15.05] %, P <.0001) than HI. A subset of allo-HSCT recipients (VZV-responders, n = 15/57, 26%) distinguished themselves from VZV-non-responders (n = 42/57, 74%; missing data, n = 3) by higher IFN-γ release (80.45 [54.3–312.8] vs. 0.22 [0.12–0.42] pg/ml, P <.0001) and T-cell proliferation (2.22 [1.18–7.56] % vs. 0.002 [0.001–0.11] %, P <.0001), suggesting recovery of VZV-specific CMI. Interestingly, VZV responders had a significant fold increase in ifn-γ gene expression, whereas ifn-γ mRNA was not detected in whole blood of VZV-non-responders (P <.0001). This study is the first to suggest that measurement of ifn-γ gene expression in 24-h-stimulated whole blood could be an accurate test of VZV-specific CMI. The routine use of this immune functional assay to guide antiviral prophylaxis at an individual level remains to be evaluated.

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“…After rapid thawing and overnight resting, PBMCs were stimulated with one µg/mL of spike (S), nucleocapsid (N) or membrane (VME1) mega pools of SARS-CoV-2 peptides from the ancestral strain, as well as the spike from variants B.1.1.7 (Alpha), B.1.351 (Beta), P.1 (Gamma), B.1.617.2 (Delta), and B.1.1.529 (Omicron) and from two other −coronaviruses (HCoV-OC43 and HCoV-HKU1) (JPT Peptide Technologies, JPT, Berlin, Germany) as described elsewhere. 24 Culture media without peptide (AIM-V® Medium (1X), Thermo Fisher Scientific, Waltham, MA, USA) and CytoStim TM (Miltenyi Biotec, MA, USA) were used as negative and positive controls respectively. Spots were quantified using CTL ImmunoSpot® SS UV Analyzer (Cellular Technology Ltd., OH, USA).…”

Section: Enzyme-linked Immunospot (Elispot) For the Quantification Of...mentioning

confidence: 99%

Differential Adaptive Immune Responses Following SARS-CoV-2 Infection in Children Compared to Adults

Nantel,

Arnold,

Bhatt

et al. 2024

Preprint

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Background : SARS-CoV-2 infection elicits distinct clinical features in children and adults. Profiling the adaptive immune response following infection in children is essential to better understand and characterize these differences. Methods : Humoral and cell-mediated immune responses from unvaccinated pediatric and adult participants were analyzed following asymptomatic or mild non-Omicron SARS-CoV-2 infection. Levels of IgG and IgA targeting spike (S), receptor-binding domain (RBD), and nucleocapsid (N) proteins of SARS-CoV-2 were measured, while neutralizing antibody (nAb) titers were assessed against three viral strains (Wuhan, Omicron BA.1 and BA.4/BA.5). Specific T-cell memory responses were investigated by quantifying interferon-gamma secreting cells after stimulation with ancestral and variant strains of SARS-CoV-2, and seasonal human coronaviruses HCoV-OC43 and HCoV-HKU1. Results : The study comprised 28 children (3 to 17 [median=10] years old) and 28 adults (19 to 62 [median=42]). At a mean time of seven months (+/- 2.8 months) after SARS-CoV-2 infection, children and adults mounted comparable antibody levels against S and RBD, as well as similar neutralization capacity. However, children displayed a weaker cellular memory response to SARS-CoV-2 than adults, with a median of 88 [28-184] spot forming units per million of PBMCs in children compared to 208 [141-340] in adults (***, P < .001). In children, the level of IFN-gamma secreting cells in response to SARS-CoV-2 corresponds to that of seasonal coronaviruses. Conclusion : Long-term memory T-cell responses to SARS-CoV-2 are enhanced in adults compared to children who demonstrate equivalent responses to SARS-CoV-2 and other HCoV.

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Development of an IFN-γ ELISpot Assay to Assess Varicella-Zoster Virus-specific Cell-mediated Immunity Following Umbilical Cord Blood Transplantation

Cited by 6 publications

References 44 publications

Herpes zoster in kidney transplant recipients: protective effect of anti-cytomegalovirus prophylaxis and natural killer cell count. A single-center cohort study

Herpes zoster in kidney transplant recipients: protective effect of anti-cytomegalovirus prophylaxis and natural killer cell count. A single-center cohort study

A Simple-to-Perform ifn-γ mRNA Gene Expression Assay on Whole Blood Accurately Appraises Varicella Zoster Virus-Specific Cell-Mediated Immunity After Allogeneic Hematopoietic Stem Cell Transplantation

Differential Adaptive Immune Responses Following SARS-CoV-2 Infection in Children Compared to Adults

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