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Distribution of MHC class II antigens in feline tissues and peripheral blood
B A Rideout1, P F Moore, N C Pedersen
1Department of Medicine, School of Veterinary Medicine, University of California, Davis.
Insights
A novel antibody, 43.2H2, targets a feline immunodeficiency virus complex on lymphocytes. This antibody shows broad reactivity across feline lymphocyte subsets, including CD4+, CD8+, and B cells, and monocytes.
Area of Science:
- Immunology
- Virology
- Cell Biology
Background:
- Feline immunodeficiency virus (FIV) is a significant pathogen in cats.
- Understanding FIV-host interactions requires specific reagents for studying viral components and host cell responses.
- Monoclonal antibodies are crucial tools for identifying viral antigens and characterizing cellular targets.
Purpose of the Study:
- To characterize a new monoclonal antibody (43.2H2) raised against FIV.
- To determine the cellular targets and reactivity patterns of antibody 43.2H2 in feline peripheral blood lymphocytes and tissues.
- To investigate the expression of the target antigen on different feline lymphocyte subsets and monocytes.
Main Methods:
- Generation of a monoclonal antibody (43.2H2) against gradient-purified FIV.
- Immunoperoxidase staining of feline tissues.
- Flow cytometric analysis of feline peripheral blood lymphocytes.
- Two-color flow cytometry to analyze reactivity on lymphocyte subsets (CD4+, CD8+, B cells) and monocytes.
Main Results:
- Antibody 43.2H2 recognizes a bimolecular complex (27-29 kD and 32-35 kD subunits) on feline lymphocytes.
- Immunoperoxidase staining showed reactivity in T-cell zones of spleen and lymph nodes, differing from human/mouse patterns.
- Flow cytometry revealed high reactivity with peripheral blood lymphocytes (88.97%), CD4+ cells (88.92%), CD8+ cells (85.99%), B lymphocytes (99.47%), and monocytes (95.06%).
- Granulocytes were negative for 43.2H2 reactivity.
Conclusions:
- Antibody 43.2H2 is a valuable reagent for detecting an FIV-associated antigen complex on feline immune cells.
- The antibody demonstrates broad reactivity with major lymphocyte populations and monocytes, suggesting a potential role in FIV pathogenesis or immune response.
- The distinct tissue staining pattern warrants further investigation into the antigen's distribution and function in feline immunity.
Abstract:
A new monoclonal antibody raised against gradient-purified feline immunodeficiency virus was found to recognize a bimolecular complex, comprising 27-29 kD and 32-35 kD subunits, on feline peripheral blood lymphocytes. Immunoperoxidase staining of feline tissues with this antibody, designated 43.2H2, demonstrated a reactivity pattern similar to that described for MHC II antigens of the dog, horse, and pig, but differed from human and mouse in having staining of T-cell zones in spleen and lymph nodes. Flow cytometric analysis revealed that 42.3H2 reacted with 88.97% +/- 16.00% of feline peripheral blood lymphocytes (n = 20). This high level of reactivity was found to be consistent by repeated sampling over a 4-month period. Two-color flow cytometric analysis was used to determined the reactivity pattern on lymphocyte subsets: 88.92% +/- 7.30% of CD4+ lymphocytes were 42.3H2-positive, while 85.99% +/- 11.46% of CD8+ cells were positive (n = 11 for both). B lymphocytes had the highest reactivity (99.47% +/- 0.45; n = 9) and also had the highest fluorescence intensity. By gating based on light scatter properties, 95.06% +/- 7.35% of monocytes were 42.3H2-reactive (n = 18), while granulocytes were negative.
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