ELISPOT assay for detection of peptide specific Interferon-γ secreting cells in rhesus macaques

A dual color ELISPOT method for the simultaneous detection of IL-2 and IFN-γ HIV-specific immune responses

“…Nevertheless, with an input of 200 000 cells/well in comprehensive screening for HIV-specific responses to all expressed HIV genes using a peptide pool matrix approach (Addo et al, 2003;Peretz et al, 2005), between 20-25 million PBMCs would be needed for the initial screen. In addition, the within-and between-assay variations were both within range reported by others for single color ELISPOT assay (Kumar et al, 2001;Mwau et al, 2002b). Moreover, depletion experiments demonstrated that although the majority of responses detected were CD8+ T cell dependent, we were also able to detect a response by CD4+ T cell secreting IL-2 alone.…”

Gamma Interferon Responses toPlasmodium falciparumLiver-Stage Antigen 1 and Thrombospondin-Related Adhesive Protein and Their Relationship to Age, Transmission Intensity, and Protection against Malaria

“…Numerous recent animal (24,38) and human (10,11,13,37,44) studies have used ELISPOT testing for IFN-␥ as their primary measure of immunity. However, in human populations it has not been established whether the level of IFN-␥ production (as measured by ELISA) or the number of mononuclear cells producing IFN-␥ (as measured by ELISPOT) is a better correlate of protection against infection and disease.…”

Protection of Rhesus Monkeys against Infection with Minimally Pathogenic Simian-Human Immunodeficiency Virus: Correlations with Neutralizing Antibodies and Cytotoxic T Cells

“…Cellular immune responses were monitored by measuring gamma interferon (IFN-␥) secretion of monkey PBMCs stimulated with SIV Gag peptide pools in an enzyme-linked immunospot (ELISPOT) assay (35 were coated with anti-human/monkey IFN-␥ antibody (MAb GZ-4; Mabtech AB, Nacka Strand, Sweden). Freshly isolated PBMCs from each monkey were added to the coated plates at a density of 2 ϫ 10 5 cells per well.…”