Cerny J, Fistel S H, Hensgen P A
Int J Cancer. 1976 Aug 15;18(2):176-88. doi: 10.1002/ijc.2910180207.
A new technique for infection of mature lymphocytes with murine leukemia virus (Friend) MuLV-F) is described. Spleen cells for normal, non-infected donors were placed into diffusion chambers (constructed with 0.22 mum por size Millipore filters) which were then implanted into the peritoneal cavities of normal syngeneic recipient mice. The cells were infected with an injection of MuLV-F into the peritoneal cavity and, in some instances, also by placing virus into the chambers. Cells were recovered by treating the chamber content with elastase and collagenase. The infection was determined in two ways: (1) cells with replicating MuLV were enumerated as infection centers (IC) on S+L- indicator cells; and (2) virus-related cell membrane antigen (MA) was detected by immunofluorescence. Cells recovered from chambers after 2-3 weeks of culture represented about 10% of the original inoculum; viability was approximately 90%. The number of IC in MuLV-F-infected chambers was about 10 times higher than obtained by infection and cultivation of spleen cells in vitro. The kinetics of IC and MA in chamber-cultured. MuLV-F-infected spleen cells was similar to that in the spleen of infected mice during the first 10 days after infection. Later on, the process of infection within the chambers slowed down, reaching about 50% MA-positive and about 10% IC-positive cells, whereas the number of both IC- and MA-positive cells in the spleen reached 80% or more. The infection of splenic lymphocytes in diffusion chambers occurred equally well when chambers were implanted into: (1) syngeneic, virus susceptible hosts; (2) syngeneic, lethally irradiated hosts; and (3) allogeneic, virus-resistant hosts, suggesting that the process within the chamber is independent of MuLV replication in the tissues of the chamber-bearing mouse. The diffusion chamber technique seems to provide an environment in which various types of isolated lymphocytes of different mouse strains can interact with MuLV almost as efficiently as in vivo.
本文描述了一种用鼠白血病病毒（Friend）（MuLV-F）感染成熟淋巴细胞的新技术。将来自正常、未感染供体的脾细胞置于扩散小室（由孔径为0.22μm的密理博滤膜构建）中，然后将这些小室植入同基因正常受体小鼠的腹腔。通过向腹腔注射MuLV-F感染细胞，在某些情况下，也通过将病毒放入小室来感染细胞。通过用弹性蛋白酶和胶原酶处理小室内容物来回收细胞。感染通过两种方式确定：（1）将具有复制性MuLV的细胞作为感染中心（IC）在S+L-指示细胞上计数；（2）通过免疫荧光检测病毒相关细胞膜抗原（MA）。培养2-3周后从小室中回收的细胞约占原始接种物的10%；活力约为90%。MuLV-F感染小室中的IC数量比体外感染和培养脾细胞获得的数量高约10倍。小室培养的MuLV-F感染脾细胞中IC和MA的动力学在感染后的前10天与感染小鼠脾脏中的相似。后来，小室内的感染过程减缓，达到约50%的MA阳性和约10%的IC阳性细胞，而脾脏中IC和MA阳性细胞的数量达到80%或更多。当将扩散小室植入以下小鼠时，脾淋巴细胞在小室内的感染效果相同：（1）同基因、病毒易感宿主；（2）同基因、经致死剂量照射的宿主；（3）异基因、病毒抗性宿主，这表明小室内的过程独立于携带小室小鼠组织中的MuLV复制。扩散小室技术似乎提供了一种环境，在这种环境中，不同小鼠品系的各种类型的分离淋巴细胞与MuLV相互作用的效率几乎与体内相同。
