Analysis of bovine B cell reactive monoclonal antibodies.

Monoclonal antibodies (mAbs) submitted to the workshop B cell panel were screened for reactivity with bovine surface immunoglobulin positive (SIg+) cells from tonsil, mesenteric lymph nodes (MLN), ileal Peyer's patches (IPP), thymus and peripheral blood by fluorescence activated cell sorter (FACS) analysis. All mAbs within B cell panel reacted with B cells except the negative control mAb VPM61. Although most mAbs stained the majority of B cells from different tissues, 14 mAbs did not stain B cells from IPP. Detailed FACS analysis of the 24 mAbs in preliminary clusters PC17, PC28, PC29, PC30, PC35, PC41 and PC43 showed all preliminary clusters (PC) to contain mAbs with similar and variable specificities. This absence of clear-cut clusters therefore did not permit a simple classification of B cell surface antigens via the B cell mAbs and suggests considerable complexity and variability of the B cell surface.

Identification of bovine B cell reactive and B cell specific monoclonal antibodies.

All monoclonal antibodies (mAbs) submitted to the workshop panel were screened for reactivity with bovine surface immunoglobulin (sIg)+ cells (gated small dense lymphocytes from peripheral blood) by fluorescence activated cell sorter (FACS) analysis. Eighteen temporary clusters--TCs 1-12, 15, 16, 18, 19, 25 and 26--contained mAbs reactive with sIg+ cells. mAb BAS21A (unclustered) and CC92 (TC25) were also reactive with sIg+ cells. Further FACS analysis with B cells from peripheral blood lymphocytes and mesenteric lymph nodes, and B and T lymphoma cell lines, indicated that the majority of mAbs within TCs 2, 4, 15, 18 and 26 reacted specifically with bovine B cells. Bovine B cell specific mAbs within these clusters were TH14B, IL-A55, CACT101A, MUC76A from TC4, VPM30, GC65A, CACT65A from TC15, IL-A58, CC56, CC70, IL-A65 from TC18, and CC57 and 26A9 from TC26. Three mAbs--IL-A65, CC70, and BAQ15A--within TC18 defined WC3; mAbs TD9 and CC56 may also be related to WC3.