McColl K A, Calnek B W, Harris W V, Schat K A, Lee L F
Department of Avian and Aquatic Animal Medicine, New York State College of Veterinary Medicine, Cornell University, Ithaca 14853.
J Natl Cancer Inst. 1987 Nov;79(5):991-1000.
Various avian tumor cell lines and normal spleen cells from 3 genetic strains of specific-pathogen-free (SPF) chickens were examined for expression of Marek's disease (MD) tumor-associated surface antigen (MATSA). Two anti-MATSA monoclonal antibodies (RPH 6 and EB 29) and a rabbit anti-MATSA antiserum were used in indirect fluorescent antibody tests, and cells were examined by fluorescence microscopy and with a fluorescence-activated cell sorter (FACS). Less than 5% MATSA-positive cells were observed in 2 non-MD tumor cell lines (LSCC-RP 9 and RECC-CU 60) with RPH 6, but 7-82% positive cells were observed with EB 29 or the rabbit antiserum. Five MD tumor cell lines (MDCC-CU 2, -CU 14, -CU 25, -CU 32, and -CU 41) had 12-72% positive cells detected with one or both monoclonals and 31-99% positive cells detected with the rabbit antiserum. Over 90% of cells in all MD lines were la and T3 positive, while values for the same parameters in LSCC-RP 9 were 100 and 3% and for RECC-CU 60, 48 and 51%, respectively. Evidence for cell-cycle-dependent expression of MATSA on MDCC-CU 2 was obtained from cell sorting experiments with the FACS and from examination of the MATSA-staining characteristics of 3 clones derived from the parent culture. Less than 5% MATSA-positive cells were observed in uncultured spleen cells from SPF chickens or in spleen cells stimulated for 48 hours with concanavalin A or phytohemagglutinin-M. However, with one exception, 10-53% of normal spleen cells were MATSA positive with RPH 6, after stimulation by mitogen for 24 or 48 hours followed by maintenance in conditioned medium (CM) for various times or after culture directly in CM for 3 days. More limited experiments with rabbit anti-MATSA antiserum yielded 55-85% MATSA-positive cells. From 60 to 97% of these MD virus-free, MATSA-positive cells were la-positive; and, in 2 cases, 89 and 90% were T3 positive.
对来自3个无特定病原体（SPF）鸡遗传品系的多种禽肿瘤细胞系和正常脾细胞进行了马立克氏病（MD）肿瘤相关表面抗原（MATSA）表达的检测。在间接荧光抗体试验中使用了两种抗MATSA单克隆抗体（RPH 6和EB 29）以及一种兔抗MATSA抗血清，并通过荧光显微镜和荧光激活细胞分选仪（FACS）对细胞进行检测。在用RPH 6检测时，在2种非MD肿瘤细胞系（LSCC-RP 9和RECC-CU 60）中观察到少于5%的MATSA阳性细胞，但在用EB 29或兔抗血清检测时，观察到7%-82%的阳性细胞。5种MD肿瘤细胞系（MDCC-CU 2、-CU 14、-CU 25、-CU 32和-CU 41）用一种或两种单克隆抗体检测到12%-72%的阳性细胞，用兔抗血清检测到31%-99%的阳性细胞。所有MD细胞系中超过90%的细胞la和T3呈阳性，而LSCC-RP 9中相同参数的值分别为100%和3%，RECC-CU 60中分别为48%和51%。通过FACS进行细胞分选实验以及对源自亲代培养物的3个克隆的MATSA染色特征进行检查，获得了MDCC-CU 2上MATSA细胞周期依赖性表达的证据。在SPF鸡的未培养脾细胞中，或在用伴刀豆球蛋白A或植物血凝素-M刺激48小时的脾细胞中，观察到少于5%的MATSA阳性细胞。然而，除一个例外，在用丝裂原刺激24或48小时后，在条件培养基（CM）中培养不同时间或直接在CM中培养3天后，10%-53%的正常脾细胞用RPH 6检测呈MATSA阳性。用兔抗MATSA抗血清进行的更有限的实验产生了55%-85%的MATSA阳性细胞。这些无MD病毒、MATSA阳性细胞中有60%-97%为la阳性；在2例中，89%和90%为T3阳性。
