Correlation between postoperative prognosis in gastrointestinal cancer patients and blastformation rate of lymphocytes

Gastrointestinal cancer patients were followed up for up to 30 months postoperatively and their clinical status related to a parameter of nonspecific immunity, the blastformation rate of peripheral blood lymphocytes against phytohemagglutinin. By the fourth postoperative week, the blastformation rate had recovered from the effect of the operation. In patients who had undergone curative resection, the postoperative level rose to exceed the preoperative level, whereas whereas in those in whom resection had not been possible, the blastformation rate failed to show this rise by the fourth week, and continued at the decreased immediate postoperative level. Results for long-term follow-up (30 months postoperatively) showed that the blastformation rate continued at high levels (almost all over 40%) in cases of curative resection without recurrence, but remained low (under 40%) in those in which the tumor could not be removed. The 40% level of the blastformation rate test thus correlated well with the prognosis. The blastformation rate, therefore, proved a very good parameter for following the pre-and post-operative clinical course of gastrointestinal cancer patients.</p

Tissue typing by mixed culture of lymphocytes. 3. Demonstration of correlation between the rate of blastformation in mixed lymphocyte culture of dog peripheral blood and the intensity of kidney-transplant rejection

1) It has been found that the peripheral blood of hybrid adult dogs contains about 29.4 % of lymphocytes in average. When such a blood is passed through the glass wool column the leucocytes containing 68-90 % (81.6% in average) lymphocytes are obtained. 2) In the single culture of such lymphocytes alone and mixed culture both live lymphocytes and sonicated lymphocytes in the presence of 1% (vIv) PHA, the peak of the blastformation of lymphocytes is observed at culture hour 72. 3) In the abscence of PHA both single culture and mixed culture of lymphocytes show hardly any blastformation. 4) In the mixed culture of live lymphocytes with homogenate of sonicated lymphocytes with addition of 1% (vIv) PHA the rate of blast. formation observable at culture hour 72 and the rejection of kidney trans· plant 7 days after its transplantation of hybrid adult dogs show a direct correlation, demonstrating that the mixed lymphocyte reaction reflects accurately the difference in donor histocompatibility antigens against the recipient.</p

Tissue typing by mixed culture of lymphocytes. I. Demonstration of intracellular localization of transplantation antigen and H-2 antigen differences by the mixed cultures with addition of whole homogenate or ultracentrifuged fractions of mouse lymph-node cells

1. The cells used in the present experiments were lymph-node cells from inbred mice, and over 98 % cells were proven to be small lympho-cytes. Therefore, those cells that have undergone blastformation are all those derived from small lymphocytes. 2. When homogenate of one cell group is cultured with live cells of the other pairing group, there occurs blastformation. In the presence of PHA, such a blastformation becomes more marked. 3. The optimal concentration of PHA (phytohemagglutinin)-M added to the mixed culture is found to be 1% (v/v). 4. The maximum rate of blastformation in the mixed culture is observed at the culture hour 48, being much faster than in the mixed culture between two live cell groups. 5. In the mixed cultures between subcellular fractions prepared from cell homogenate by centrifugation and live cells, the transplantation antigenic potential (histocompatibility antigenic potential) is seen in the mitochondrial and the microsomal fractions, especially marked in the latter. 6. In the observations carried out by various combinations of these inbred mice, it has been demonstrated that the rate of blastformation induced by the addition of cell homogenate or sediment fractions prepared from the homogenate reflects quite accurately the differences in H-2 antigens.</p

Tissue typing by mixed culture of lymphocytes. II. Demonstration of H-2 antigen differences by mixed cultures with addition of subcellular fractions prepared from homogenate of lymph-node cells destroyed by supersonication

1. It has been found that mouse lymph.node cells, even destroyed by sonication with 20 KC supersonicator, maintain sufficient antigenicity both in vitro and in vivo. 2. When such sonicated cell homogenate is cultured with live lymph-node cells, there can be observed blastformation and the peak of the rate of the blastformation is seen at culture hour 48. 3. When PHA (phytohemagglutinin)-M is added to such mixed cultures, the blastformation is enhanced. 4. When mixed cultures of mouse lymph-node cells are conducted by using such one-way stimulation method in various combinations, the rate of blastformation can tell quite accurately the differences in H-2 antigens of mice. 5. In the experiment using F1 hybrid mice and the parents, it has been demonstrated that the rate of blastformation in mixed cultures of the present experiments shows a direct correlation to the rate of blast formation in mixed cultures of live lymph node cells, whlie it is an inverse proportion to the survival time of the skin transplant. 6. Differences in the transplanation antigens said to be located on sex chromosomes cannot be distinguished by this one.way stimulation method.</p

Reactivating effect of levamisole on cell-mediated immunity in gastrointestinal cancer patients

Cell-mediated immunity was studied in 23 cases of advanced gastrointestinal cancer. The patients received levamisole at 150 mg/day for three consecutive days each week for four weeks. In cases at the terminal stage of gastrointestinal cancer, the blastformation rate of peripheral blood lymphocytes against phytohemagglutinin (PHA) after the administration of levamisole showed a slight increase, but cases with blastformation rates over 40% increased markedly three or four weeks after the initial administration of levamisole. The peripheral blood lymphocyte count showed little change in these cases.</p

Immunotherapy of gastric cancer with levamisole

Ninety-nine gastric cancer patients initially received levamisole at a daily dose of 150 mg for three consecutive days before operation. This therapy was repeated fortnightly (3-day administration followed by 11-day withdrawal period) for more than one month as long as possible and the survival rate up to 18 months was compared with thas of control patients. The 18 month survival rate of advanced Stage IV patients was significantly higher in patients receiving levamisole than that of control patients. The effects of levamisole in cases of advanced cancer have been discussed in relation to the literature available.</p

Cancer immunotherapy with levamisole

Levamisole, an agent acting upon depressed cellular immunity, enhancing and normalizing it and consequently showing antitumor activity in the cancer-bearing body, was administered to patients with gastrointestinal cancer at a daily dose of 150 mg for three consecutive days every other week, starting as a rule, three days before operation. The patients were evaluated for survival. Of the 143 patients (66 with curative resection, 40 with noncurative resection and 37 without resection) who received levamisole therapy for one month or more, 57 survived postoperatively six months and of 44 treated 37 survived one year. In this study, 185 patients with gastrointestinal cancer were used for comparison purposes. The six-month survival rate was 100% (23/23) in the levamisole treated group and 95.3% (102/107) in the control group after curative resection (p greater than 0.5), 100% (23/23) and 90.5% (49/54) after noncurative resection (p less than 0.01), and 72.5% (8/11) and 33.3% (9/24), respectively, in non-resectable patients (p less than 0.01). The one-year survival rate was 100% (21/21) and 95.3% (102/107) after curative resection (p greater than 0.5), 77.8% (14/18) and 59.3% (32/54) after noncurative resection (0.05 less than p less than 0.1), and 40% (2/5) and 8.3% (2/24) in non-resectable patients (0.05 less than p than 0.1) in the levamisole group and in the control group, respectively. The difference in survival in survival rates between levamisole-treated and control groups was most prominent in the non-resectable patients followed by those undergoing noncurative resection and curative resection.</p

Preoperative blastformation rate in gastrointestinal cancer patients

The rate of blastformation of peripheral blood lymphocytes in response to stimulation by phytohemagglutinin (PHA) was assessed preoperatively in 393 patients with gastrointestinal cancer. The series consisted of 291 cases of gastric cancer and 102 cases of colon cancer, all patients being under 70 years of age. The blastformation rate was related to the stage of cancer ground at operation. Preoperative blastformation rates for both colon cancer and gastric cancer decreased as the cancer progressed. With Stage I gastric cancer 81.4% of those that underwent curative resection had preoperative blastformation rates greater than 40%. However, the number of those with blastformation rates over 40% decreased markedly in the curative cases of gastric cancer Stage II to stage IV. Eighty three percent of cases that underwent curative resection with colon cancer, including advanced cancer, had preoperative blastformation rates of over 40%. These results indicated that the correlation of the preoperative blastformation rate with success of curative resection better for colon cancer than for gastric cancer.</p

Correlation between morphological blastformation rate and functional 3H-thymidine uptake in mixed lymphocyte culture in the presence of PHA

By dividing at random 14 normal persons into 7 pairs of two individuals each, lymphocytes were isolated from their peripheral blood and taking one of the pairs as stimulating cells or antigens and the others as responding cells, mixed lymphocyte culture (MLC) was carried out. As for the method of MLC we used our MLC method of unidirectional mixed culture with a small amount of lymphocytes in additition of 1% (v /v) PHA.M and cultured for three days, and a widely used conventional method in which 3H.thymidine uptake was the parameter of the blastformation rate and cultured for seven days. In comparing the results of these two groups of MLC the data in six experiments out of the seven coincided. Namely, with 5x 104 cells each of stimulating cell group and responding cell group, it is possible to achieve satisfactory MLC, the culture can be done only for three days without requiring any special technique and the results can be readily evaluated. Therefore, MLC by our simple method would yield satisfactory results in clinics.</p

Identification and prognostic implications of tumor infiltrating lymphocytes--a review.

Cancer patients who have many tumor infiltrating lymphocytes (TIL) tend to have better prognoses. A relationship between prognosis and TIL or regional lymph node response is present in several malignant diseases. TIL are mainly T lymphocytes, as ascertained by immunological methods. Results of studies on T-lymphocyte subsets comprising TIL using monoclonal antibodies (OKT series and Leu series) are summarized in this review.</p