tetano
Editor, Senior Moderator
J Immunol Methods. 2013 Sep 18. pii: S0022-1759(13)00247-0. doi: 10.1016/j.jim.2013.09.007. [Epub ahead of print]
Granzyme B ELISPOT assay to measure influenza-specific cellular immunity.
Salk HM, Haralambieva IH, Ovsyannikova IG, Goergen KM, Poland GA.
Source
Mayo Vaccine Research Group, USA.
Abstract
The immunogenicity and efficacy of influenza vaccination are markedly lower in the elderly. Granzyme B (GrzB), quantified in fresh cell lysates, has been suggested to be a marker of cytotoxic T lymphocyte (CTL) response and a predictor of influenza illness among vaccinated older individuals. We have developed an influenza-specific GrzB ELISPOT assay using cryopreserved PBMCs. This method was tested on 106 healthy older subjects (ages 50-74) at baseline (Day 0) and three additional time points post-vaccination (Day 3, Day 28, Day 75) with influenza A/H1N1-containing vaccine. No significant difference was seen in GrzB response between any of the time points, although influenza-specific GrzB response appears to be elevated at all post-vaccination time points. There was no correlation between GrzB response and hemagglutination inhibition (HAI) titers, indicating no relationship between the cytolytic activity and humoral antibody levels in this cohort. Additionally, a significant negative correlation between GrzB response and age was observed. These results reveal a reduction in influenza-specific GrzB response as one ages. In conclusion, we have developed and optimized an influenza-specific ELISPOT assay for use with frozen cells to quantify the CTL-specific serine protease GrzB, as a measure of cellular immunity after influenza vaccination.
? 2013.
KEYWORDS:
Cellular, Cellular immunity, Cytotoxic, Cytotoxic T lymphocytes (CTL), ELISPOT, Enzyme-linked immunospot assay, Granzyme B, Granzymes, Human, Immunity, Influenza, Influenza virus, Killer cells, Natural, Perforin, T-lymphocytes, Viruses
PMID:
24055591
[PubMed - as supplied by publisher]
http://www.ncbi.nlm.nih.gov/pubmed/24055591
Granzyme B ELISPOT assay to measure influenza-specific cellular immunity.
Salk HM, Haralambieva IH, Ovsyannikova IG, Goergen KM, Poland GA.
Source
Mayo Vaccine Research Group, USA.
Abstract
The immunogenicity and efficacy of influenza vaccination are markedly lower in the elderly. Granzyme B (GrzB), quantified in fresh cell lysates, has been suggested to be a marker of cytotoxic T lymphocyte (CTL) response and a predictor of influenza illness among vaccinated older individuals. We have developed an influenza-specific GrzB ELISPOT assay using cryopreserved PBMCs. This method was tested on 106 healthy older subjects (ages 50-74) at baseline (Day 0) and three additional time points post-vaccination (Day 3, Day 28, Day 75) with influenza A/H1N1-containing vaccine. No significant difference was seen in GrzB response between any of the time points, although influenza-specific GrzB response appears to be elevated at all post-vaccination time points. There was no correlation between GrzB response and hemagglutination inhibition (HAI) titers, indicating no relationship between the cytolytic activity and humoral antibody levels in this cohort. Additionally, a significant negative correlation between GrzB response and age was observed. These results reveal a reduction in influenza-specific GrzB response as one ages. In conclusion, we have developed and optimized an influenza-specific ELISPOT assay for use with frozen cells to quantify the CTL-specific serine protease GrzB, as a measure of cellular immunity after influenza vaccination.
? 2013.
KEYWORDS:
Cellular, Cellular immunity, Cytotoxic, Cytotoxic T lymphocytes (CTL), ELISPOT, Enzyme-linked immunospot assay, Granzyme B, Granzymes, Human, Immunity, Influenza, Influenza virus, Killer cells, Natural, Perforin, T-lymphocytes, Viruses
PMID:
24055591
[PubMed - as supplied by publisher]
http://www.ncbi.nlm.nih.gov/pubmed/24055591