Alistair J. Cochran

RESULTS OF ADMINISTERING B.C.G. TO PATIENTS WITH MELANOMA

Abstract Ten patients with malignant melanoma were given immune stimulation by B.C.G. scarification, intratumoral B.C.G., or B.C.G. in combination with irradiated autologous tumour cells. In addition, four patients received antitumour antibody associated with chlorambucil by intravenous infusion. In six patients objective clinical improvement was observed. Complications included granulomatous hepatitis, mycobacterial pneumonia, ulceration of the site of injection of tumour cells and B.C.G., and splenomegaly. Necropsy findings included multiple granulomas in three patients, in two of whom acid/ alcohol-fast bacilli were identified. Anaphylaxis complicated infusion of antitumour antibodies and chlorambucil in one case. During B.C.G. treatment seven negative Mantoux skin tests became positive, immunoglobulin levels increased in five patients, leucocyte migration continued to be inhibited by tumour-derived materials in patients with advanced disease, and cytotoxic lymphocytes lethal to melanoma cells developed in two patients.

Thirteen new mammary tumor cell lines from different mouse strains

Abstract Seventeen mammary tumors transplantable in various strains of mice were cultured to obtain established cell lines. All but four tumors have been successfully established as monolayers, have been subcultured regularly, and produced tumors on inoculation into syngeneic mice. Histological examination showed that seven lines (SHF and SHK in C3H mice, S6C in ACA mice, S3W in ASW mice, S2Y in ABY mice, SBfnHA and SBfnHB in CBA mice) are adenocarcinomas, three (TA3Ha, TA3St in A mice and SBfnHD in CBA mice) are poorly differentiated carcinomas, and three (SHG in C3H mice, S40C in ACA mice and SBfnHC in CBA mice) are spindle celled, ‘sarcoma-like’ tumors associated with much reticulin and collagen. Electron-microscopy demonstrated that all lines, including the ‘sarcoma-like’ SHG, S40C and SBfnHC are carcinomatous. Eleven lines showed secretion vacuoles consistent with the presence of milk protein. SHG, S40C and SBfnHC are thus to be regarded as spindlecelled carcinomas. All cell lines, except SHG and SBfnHC, were sensitive to antimammary tumor virus serum. SHK, S2Y and SBfnHB showed extremely high reactivity to this antiserum. These thirteen established cultured lines promise to be useful in the further study of mouse mammary tumors.

Viral expression and immunogenicity of CBA mammary carcinomas and their hybrid lines with an L-cell derivative (A9HT)☆

Abstract Somatic cell hybrid lines were obtained by Sendai virus mediated fusion of a highly tumorigenic variant of L-cell subline A9 of C3H mouse origin (A9HT) with two CBA mouse mammary carcinomas (SBfnHA and SBfnHC) that had been inoculated into CBAT6 mice. Hybridity was confirmed by the presence of biarmed marker chromosomes from the A9HT parent, and the total chromosome number, which in both hybrids (SBfnHA/A9HT and SBfnHC/A9HT) approximated the sum of the parental chromosomes. The absence of the T6 marker in both the hybrids confirmed that fusion was not between a stroma cell from CBAT6 mice and A9HT. Both hybrids showed spindle cell carcinoma morphology. Complement dependent cytotoxicity assays revealed that SBfnHA and A9HT expressed the MTV-associated antigens including one major structural component, gp52 and the MuLV structural components gp71. and p30 on the cell-surface, while SBfnHC expressed only gp71. The hybrid line SBfnHA/A9HT lost the surface expression of MTV gp52 and MuLVp30, and there was also decreased expression of gp71 in comparison with SBfnHA. In contrast, in SBfnHC/A9HT there was increased expression of gp71 over that of SBfnHC. All parental and hybrid lines had unchanged sensitivity to the cytotoxic effect of anti-H-2 k sera. Immunization of CBA mice with heavily irradiated either syngeneic SBfnHA or hybrid SBfnHA/A9HT cells, followed by challenge with viable SBfnHA cells revealed that SBfnHA was a highly immunogenic tumor, while the hybrid line SBfnHA/A9HT decreased in the immunogenicity of the parental SBfnHA line. On the other hand, immunization of CBA mice with heavily irradiated either syngeneic SBfnHC or hybrid SBfnHC/A9HT cells, followed by challenge with viable SBfnHC cells showed that SBfnHC/A9HT slightly increased the very low immunogenicity of the parental SBfnHC. These results are consistent with the possibility that MTV-and MuLV-associated cell-surface antigens influence the immunogenicity of CBA mammary carcinomas.

The Mechanism of Tumour Cell Induced Inhibition of Human Leucocyte Migration

Abstract We investigated the mechanism of leucocyte migration inhibition by formalin-fixed tumour cells. Peripheral blood leucocytes were separated into the component populations; granulocytes, lymphocytes, lymphocyte subpopulations and monocytes and incubated for 24 hr with formalinised melanoma cells and formalinised control cells. The supernatants of these cultures were tested for their capacity to inhibit the migration in vitro of normal peripheral blood leucocytes. Inhibitory activity was generated by cocultures of melanoma cells with populations containing a substantial proportion of lymphocytes. Studies with populations enriched for T lymphocytes indicated that T cells generated the inhibitory activity although our inability totally to purify the T cells meant that a (cooperative) role for Fc receptor bearing lymphocytes including B cells could not be excluded.

Subpopulations of Lymphocytes: A Review

Human lymphocytes comprise a heterologous population of cells which can be sub-divided on functional criteria or on the basis of surface membrane markers. Although it is widely accepted that human lymphocytes are broadly divisible into thymus dependent (T) and thymus independent (B) cells, recent evidence suggests that at least in humans this distinction may not be as clear cut as previously thought. An assessment of the relative numbers of T and B cells in human peripheral blood and lymphoid tissues can be made utilizing these marker systems. This has led to a reclassification of human lymphoid diseases according to the T and B cell concept, which might be of value in characterising the immunological status in health and disease, and may allow a more rational approach to the therapy of immune deficiency states and lymphoid malignancies.