Lamina propria T cell activation: role of the costimulatory molecule CD2 and its cytoplasmic tail for the regulation of proliferation and apoptosis.
OBJECTIVE
Accumulation of T lymphocytes in the gut is a hallmark of inflammatory bowel disease probably caused by insufficient T cell apoptosis. Activated peripheral T cells, or "resting" lamina propria T lymphocytes (LPLs), are highly susceptible to apoptosis induction, e.g., using the mitogenic anti-CD2 monoclonal antibody (mAb) pair T11(2+3). It is, however, unknown how CD2-mediated LPL apoptosis is related to proliferation and whether the whole CD2 molecule is required for apoptosis induction.


METHODS
Mapping of anti-CD2 mAb was performed using erythrocyte rosetting assays and cross-blocking enzyme-linked immunosorbent assay (ELISA). Lamina propria mononuclear cells (LPMNCs) or phytohemagglutinin (PHA) blasts were stimulated with a panel of 18 anti-CD2 mAbs followed by apoptosis analysis [Annexin V expression on propidium iodide (PI)-negative cells, 4c6-diamidino-2-phenylindole x 2HCl (DAPI) staining]. Proliferation was measured by [(3)H]-thymidine incorporation. For structural analysis, EL4 cells were used which were transfected with human CD2 (wild type (WT), cytoplasmic-deficient, cytoplasmic CD28). Sorting was performed employing standard techniques


RESULTS
All three mitogenic anti-CD2 mAb pairs induced apoptosis of LPMNC and PHA blasts. Two out of four submitogenic anti-CD2 mAb, AICD2.M3, and ICRFCD2.3 lead to LPMNC proliferation but no apoptosis. Importantly, apoptosis was also detected in cytoplasmic-deficient CD2 tg or CD2/CD2/CD28 tg EL4 cells. Sorted CD45(high) huCD2 WT EL4 had higher apoptosis rates compared to WT huCD2tg EL4 cells


CONCLUSIONS
LPMNC apoptosis induction via CD2 is always associated with proliferation, although proliferation is not necessarily associated with apoptosis. The cytoplasmic tail of CD2 is not required, and CD45 appears to transmit apoptotic signals entering the T cell via CD2.
Henschke
Sven
S
Innere Medizin II, Medizinische Klinik, Universitätskliniken des Saarlandes, 66421, Homburg/Saar, Germany.
Pawlowski
Nina N
NN
Wild
Martin K
MK
Kroesen
Anton J
AJ
Zeitz
Martin
M
Hoffmann
Jörg C
JC
D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology; D017209:Apoptosis; D018801:CD2 Antigens / Q000276:immunology*; D049109:Cell Proliferation; D003593:Cytoplasm; D004797:Enzyme-Linked Immunosorbent Assay; D006801:Humans; D066298:In Vitro Techniques; D017493:Leukocyte Common Antigens / Q000276:immunology; D007963:Leukocytes, Mononuclear / Q000276:immunology; D008213:Lymphocyte Activation / Q000276:immunology*; D009092:Mucous Membrane / Q000473:pathology; D010835:Phytohemagglutinins
D000911:Antibodies, Monoclonal; D018801:CD2 Antigens; D010835:Phytohemagglutinins; D017493:Leukocyte Common Antigens; C516769:PTPRC protein, human