Assessing antigen specific HLA-DR plus antibody secreting cell (DR plus ASC) responses in whole blood in enteric infections using an ELISPOT technique

作者：Bhuiyan Taufiqur Rahman^*; Hoq Mohammad Rubel; Nishat Naoshin Sharmin; Al Mahbuba Deena; Rashu Rasheduzzaman; Islam Kamrul; Hossain Lazina; Harris Jason B; Ryan Edward T; Calderwood Stephen B; Svennerholm Ann Mari; Qadri Firdausi

来源：Microbes and Infection, 2018, 20(2): 122-129.

DOI：10.1016/j.micinf.2017.10.005

• HLA-DR
• ASCs
• Immune response
• Cholera
• ETEC
• ELISPOT

Antibody secreting cells (ASCs) generate antibodies in an antigen-specific manner as part of the adaptive immune response to infections, and these cells increase their surface expression of HLA-DR. We have studied this parameter (HLA-DR+ASC) in patients with recent diarrheal infection using immuno-magnetic cell sorting and an enzyme linked immunospot (ELISPOT) technique that requires only one milliliter of blood. We validated this approach in adult patients with cholera (n = 15) or ETEC diarrhea (n = 30) on days 2, 7 and 30 after showing clinical symptom at the International Centre for Diarrhoeal Disease Research, Bangladesh (icddr, b) hospital in Dhaka, and we compared responses to age-matched healthy controls (n = 7). We found that HLA-DRthorn ASC (DR+ASC) responses specific both for T cell-dependent (cholera toxin B subunit), and T cell-independent (lipopolysaccharide) antigens were elevated at day 7 after showing clinical cholera symptom. Similarly, DR+ASCs were elevated against both heat-labile toxin and colonization factors following ETEC infection. We observed significant correlations between antigen-specific DR+ASC responses and antigen-specific, gut homing ASC and plasma antibody responses. This study demonstrates that a simple ELISPOT procedure allows determination of antigen-specific ASC responses using a small volume of whole blood following diarrhea. This technique may be particularly useful in studying DR+ASC responses in young children and infants, either following infection or vaccination.

• 出版日期2018-2