ELISPOT assay of interferon-γ secretion for evaluating human cytomegalovirus reactivation risk in allo-HSCT recipients

J Med Virol. 2021 Nov;93(11):6301-6308. doi: 10.1002/jmv.27120. Epub 2021 Jun 9.

Abstract

Human cytomegalovirus (HCMV) is a common cause of significant morbidity and mortality in transplant recipients after allogeneic hematopoietic stem cell transplantation (allo-HSCT). We evaluated interferon-γ (IFN-γ) secretion by HCMV NLV-specific CD8+ T cells in HCMV-reactivated allo-HSCT recipients using an enzyme-linked immunospot (ELISPOT) assay at 3 months post-transplantation. Blood samples from 47 recipients were tested for HCMV DNAemia, HCMV pp65 antigenemia, and anti-HCMV immunoglobulins (IgG/IgM) over 3 months post-transplantation. Of the 47 transplant recipients, 26 were HLA-A*02 positive and 21 were HLA-A*02 negative. The results were essentially consistent between the 47 transplant recipients and the HLA-A*02-positive recipients. HCMV DNAemia was not linearly correlated with IFN-γ spot-forming cells (SFCs) counts; IFN-γ SFCs counts did not differ significantly between the HCMV DNAemia-positive and -negative groups, whereas the HCMV-DNA virus loads were inversely correlated with the IFN-γ SFCs counts. HCMV pp65 antigenemia was not linearly correlated with IFN-γ SFCs counts; IFN-γ SFCs counts in the HCMV pp65 antigenemia-positive and -negative groups were similar. More IFN-γ SFCs counts were detected in transplant recipients with high anti-HCMV-IgG antibody titers than in those with low anti-HCMV-IgG titers pre-transplantation in the 47 recipients. Anti-HCMV-IgG antibody titers were positively linearly correlated with IFN-γ SFCs counts in HLA-A*02-positive recipients. The HCMV infection indicators used to monitor HCMV reactivation had different values in transplant recipients. The use of the IFN-γ SFCs counts measured by ELISPOT to evaluate the risk of HCMV reactivation needs further study.

Keywords: ELISPOT; HLA-A*02; allogeneic hematopoietic stem cell transplantation; human cytomegalovirus; interferon-γ.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Adolescent
  • Adult
  • Antigens, Viral / immunology
  • CD8-Positive T-Lymphocytes / immunology
  • Cytomegalovirus Infections / blood
  • Cytomegalovirus Infections / diagnosis*
  • Cytomegalovirus Infections / immunology
  • Cytomegalovirus Infections / virology
  • Enzyme-Linked Immunospot Assay / methods*
  • Enzyme-Linked Immunospot Assay / standards*
  • Female
  • Hematopoietic Stem Cell Transplantation / adverse effects*
  • Humans
  • Interferon-gamma / analysis*
  • Interferon-gamma / immunology
  • Latent Infection / blood
  • Latent Infection / diagnosis*
  • Latent Infection / immunology
  • Latent Infection / virology
  • Male
  • Middle Aged
  • Transplant Recipients / statistics & numerical data*
  • Young Adult

Substances

  • Antigens, Viral
  • Interferon-gamma