AN EXTENSION OF THE51Cr-RELEASE ASSAY FOR THE ESTIMATION OF MOUSE CYTOTOXINS

Boyle, William

doi:10.1097/00007890-196809000-00002

Cited by 631 publications

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“…The erythrocytes were lysed with Tris-buffered NH4Cl (Boyle, 1968), and the remaining cell pellet was again washed x 2 in Medium 199. The cell pellet was then resuspended in 20 ml of M199 and was centrifuged at 60 g for 10 min to obtain neoplastic cells in the pellet (Robins et al, 1979).…”

Section: Methodsmentioning

confidence: 99%

Differential immune reactivity of tumour-intrinsic and peripheral-blood lymphocytes against autoplastic colorectal carcinoma cells

Hutchinson¹,

Heinemann²,

Symes³

et al. 1981

Br J Cancer

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Summary.-Peripheral blood lymphocytes (PBL) were obtained from 13 patients and tumour-intrinsic lymphocytes (TIL) from 20 patients with colorectal cancer. The PBL were separated on a Ficoll-Isopaque gradient and the TIL by digestion of the tumour with collagenase-DNase. Both PBL and TIL were passed through nylonwool columns and the eluted cells were co-cultured for 2 h with 51Cr-labelled tumour cells from the same patient. If patients in whom spontaneous 51Cr release from the tumour cells was greater than 3300 were excluded, PBL showed cytotoxicity for the autoplastic tumour cells in 5/10 cases and TIL in 3/10 cases (NS). In 12 cases the cytotoxicity of the TIL was compared with that for TIL from the same tumour after the lymphocytes had been washed a further 6 times in Medium 199. Three effector: target (E/T) ratios, 5:1, 10:1 and 20:1, were used. The proportion of effector populations showing cytotoxicity was 2/12 for unwashed TIL and 9/12 for washed TIL (P<0.006). At the 5:1 E/T ratio the level of cytotoxicity was not significantly greater for washed TIL, but at the 10:1 ratio washed TIL showed significantly more cytotoxicity (P <0-025). At the 20:1 E/T ratio, a comparison was possible in 15 cases and the washed TIL again showed greater cytotoxicity (P<0-001).

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Section: Methodsmentioning

confidence: 99%

Differential immune reactivity of tumour-intrinsic and peripheral-blood lymphocytes against autoplastic colorectal carcinoma cells

Hutchinson¹,

Heinemann²,

Symes³

et al. 1981

Br J Cancer

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“…Suspensions of spleen cells were treated with Tris-ammonium chloride to eliminate erythrocytes (12), and labeled with 51Cr as described (13). For the cytotoxicity assay, 2.5 × 105 labeled spleen cells (50 /xl) were incubated with 50/xl antiserum (predetermined optimal dilution in phosphate-buffered saline), or with normal mouse serum (NMS t, control) for 30 min on ice, washed once, resuspended in 100 #1 freshly-thawed rabbit serum (complement source; predetermined optimal dilution), and incubated further for 30 min at 37"C. A single pool of rabbit serum selected for low natural cytotoxicity and high complement activity was used throughout.…”

mentioning

confidence: 99%

Disproportion in T-cell subpopulations in immunodeficient mutant hr/hr mice.

Reske‐Kunz¹,

Scheid²,

Boyse³

1979

The Journal of Experimental Medicine

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Mice of the HRS strain, which carry the mutant gene hr, were examined for abnormalities in representation of the three T-cell sets Ly1, Ly23, and Ly123 in the spleen. The salient feature of hr/hr mice, which are immunologically deficient, in comparison with +/hr segregants, was a gross disproportion in numbers of cells belonging to the Ly1 and Ly123 sets, at the age of 3--3.5 mo. At this age, Ly123 cells of hr/hr spleen outnumbered Ly1 cells by 2:1, whereas in +/hr spleens Ly123 cells were outnumbered by approximately 1:2. Cells from pooled lymph nodes of hr/hr mice did not show a correspondingly gross disporprotion of Ly1 and Ly123 cells. Total counts of splenic T cells, and of B cells, were not significantly different in hr/hr and +/hr mice.

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“…Red cells in the spleen preparations were lysed according to the method of Boyle (1968), using Tris buffered ammonium chloride. Before inoculation of cells into animals, the cells were suspended in PBS with 0-1% bovine serum albumin (BSA).…”

Section: Methodsmentioning

confidence: 99%

Immunity to Murine Sarcoma Virus Induced Tumours. III. Analysis of the Cell Populations Involved in Protection from Lethal Tumour Progression of Sublethally Irradiated, MSV Inoculated, Mice

Gorczynski¹,

Norbury²

1974

Br J Cancer

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Summary.-A comparison was made between the cells responsible for demonstrable activity against MSV antigens, using both in vivo and in vitro assays. Similar cells (in terms of size and sensitivity to anti-theta serum) were detected in both assays. However, while lymphoid cells from animals at all stages post-MSV infection were active in protecting irradiated mice from the lethal effect of induction of MSV sarcomata, cells from animals at early stages post-MSV infection (when the tumour was in a progressive phase of growth) were not active in the in vitro assay. By manipulation of the in vivo assay conditions a situation was observed in which cells from " progressor animals " were able to suppress both the in vitro and in vivo activity of regressor lymphoid cells. The potential physiological role of this cell type is discussed.

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AN EXTENSION OF THE51Cr-RELEASE ASSAY FOR THE ESTIMATION OF MOUSE CYTOTOXINS

Cited by 631 publications

References 0 publications

Differential immune reactivity of tumour-intrinsic and peripheral-blood lymphocytes against autoplastic colorectal carcinoma cells

Differential immune reactivity of tumour-intrinsic and peripheral-blood lymphocytes against autoplastic colorectal carcinoma cells

Disproportion in T-cell subpopulations in immunodeficient mutant hr/hr mice.

Immunity to Murine Sarcoma Virus Induced Tumours. III. Analysis of the Cell Populations Involved in Protection from Lethal Tumour Progression of Sublethally Irradiated, MSV Inoculated, Mice

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