SAP and SLAM expression in anti-CD3 activated lymphocytes correlates with cytotoxic activity.
Signalling lymphocyte activation molecule (SLAM)-associated protein (SAP) is a small protein that is mutant in humans with X-linked lymphoproliferative (XLP) disease. Patients with XLP disease are affected by fatal EBV infection and malignant B-cell lymphomas. The increased risk for B-cell lymphomas is suggested to result from impaired immunosurveillance of B-cell proliferation by T cells. In this study, we investigated the role of SLAM and SAP in activation of effector cells with cytotoxic activity, cytokine-induced killer (CIK) cells, which are generated by non-specific stimulation of the TCR and addition of exogenous IL-2. Agonistic TCR activation 1 day after preparation (day +1) resulted in cell activation, with a peak of SLAM on day +6 visible at both the protein and mRNA level as well as membrane detectable SLAM. This increase in SLAM expression correlated significantly with SAP expression at the mRNA level as well as at the protein level. Cytotoxic activity peaked 1 day after the observed SAP and SLAM peaks. At that point in time, IL-10 secretion, which was high during the early days of culture, decreased. In conclusion, activation of peripheral blood cells with agonistic anti-CD3 antibody and exogenous IL-2, as used for generation of CIK cells, results in significant SLAM and SAP activation 5 days after TCR stimulation. This peak correlates with cytotoxic activity against tumour cells. Expression of SLAM and SAP seems to be important in the activation of cytotoxic effector cells.
Mehrle
Stefan
S
Department of Surgery, University of Heidelberg, Heidelberg, Germany.
Frank
Susanne
S
Schmidt
Jan
J
Schmidt-Wolf
Ingo Gh
IG
Märten
Angela
A
D000906:Antibodies / Q000494:pharmacology; D015703:Antigens, CD; D017252:CD3 Complex / Q000276:immunology; D045744:Cell Line, Tumor; D003602:Cytotoxicity, Immunologic; D006023:Glycoproteins / Q000032:analysis* / Q000235:genetics / Q000502:physiology; D006801:Humans; D007136:Immunoglobulins / Q000032:analysis* / Q000235:genetics / Q000502:physiology; D016130:Immunophenotyping; D016753:Interleukin-10 / Q000378:metabolism; D047908:Intracellular Signaling Peptides and Proteins / Q000032:analysis* / Q000235:genetics / Q000502:physiology; D015979:Killer Cells, Lymphokine-Activated; D008213:Lymphocyte Activation; D008214:Lymphocytes / Q000737:chemistry; D012333:RNA, Messenger / Q000032:analysis; D011956:Receptors, Cell Surface; D000071179:Signaling Lymphocytic Activation Molecule Associated Protein; D000071177:Signaling Lymphocytic Activation Molecule Family Member 1; D013997:Time Factors
D000906:Antibodies; D015703:Antigens, CD; D017252:CD3 Complex; D006023:Glycoproteins; D007136:Immunoglobulins; D047908:Intracellular Signaling Peptides and Proteins; D012333:RNA, Messenger; D011956:Receptors, Cell Surface; C114953:SH2D1A protein, human; D000071179:Signaling Lymphocytic Activation Molecule Associated Protein; D016753:Interleukin-10; D000071177:Signaling Lymphocytic Activation Molecule Family Member 1