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-v

“…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 β−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: Methodsmentioning

confidence: 99%

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

Nantel,

Arnold,

Bhatt

et al. 2024

Preprint

0

View full text Add to dashboard Cite

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.

show abstract

“…PBMC and CBMC were used in an IFN-γ ELISpot assay adapted for ZIKV, as previously described [16]. Briefly, pools of 48 to 166 different overlapping peptides based on the sequences of multiple strains of the ZIKV capsid (C), envelope (E), non-structural protein-1 (NS-1), and matrix protein (M) were obtained from a commercial supplier (JPT Peptide Technologies GmbH, Berlin, Germany) and were reconstituted according to the manufacturer's instructions.…”

Section: Ifn-γ Elispotmentioning

confidence: 99%

Zika virus infection during pregnancy and vertical transmission: case reports and peptide-specific cell-mediated immune responses

Hindle,

Depatureaux,

Fortin-Dion

et al. 2024

Arch Virol

0

View full text Add to dashboard Cite

Zika virus (ZIKV) infection in pregnant women is associated with birth defects which are more prevalent and severe the earlier in pregnancy the infection occurs. Pregnant women at risk of possible ZIKV exposure (n = 154) were screened using ELISA for ZIKV IgM and IgG. Plaque reduction neutralization tests (PRNT) confirmed ZIKV infection in 9 cases (5.84%). Two cases of vertical ZIKV transmission were confirmed by PCR. One infant showed no signs of birth defects and had a normal developmental profile despite first trimester maternal infection. In the second case, pregnancy was terminated. Production of interferon-γ (IFN-γ) by peripheral blood mononuclear cells obtained from pregnant women and umbilical cord blood was measured using enzyme-linked immunospot assay (ELISpot) following stimulation with panels of synthetic peptides derived from the sequence of ZIKV proteins. This analysis revealed that among all peptide pools tested, those derived from the ZIKV envelope protein generated the strongest IFN-γ response.

show abstract

Zika virus infection during pregnancy and vertical transmission: case reports and peptide-specific cellmediated immune responses.

Soudeyns,

Hindle,

Depatureaux

et al. 2023

Preprint

Self Cite

0

View full text Add to dashboard Cite

Zika virus (ZIKV) infection in pregnant women is associated with birth defects which are more prevalent and severe the earlier in pregnancy the infection occurs. Pregnant women at risk of possible ZIKV exposure (n = 154) were screened using ELISA for ZIKV IgM and IgG. Plaque reduction neutralization tests (PRNT) confirmed ZIKV infection in 9 cases (5.84%). Two cases of vertical ZIKV transmission were confirmed by PCR. One infant showed no signs of birth defects and had a normal developmental profile despite first trimester maternal infection. In the second case, pregnancy was terminated. Production of interferon-γ (IFN-γ) by peripheral blood mononuclear cells obtained from pregnant women and umbilical cord blood was measured using enzyme-linked immunospot assay (ELISpot) following stimulation with panels of synthetic peptides derived from the sequence of ZIKV proteins. This analysis revealed that among all peptide pools tested, those derived from the ZIKV envelope protein generated the strongest IFN-γ response.

show abstract

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

Cited by 3 publications

References 0 publications

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

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

Zika virus infection during pregnancy and vertical transmission: case reports and peptide-specific cell-mediated immune responses

Zika virus infection during pregnancy and vertical transmission: case reports and peptide-specific cellmediated immune responses.

Contact Info

Product

Resources

About