Augmentation by serum-induced helper cells of T cell-mediated cytotoxic response against tumor associated antigens and alloantigens.
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Biomedical subjects
Publications and source records attributed to D Rodrigues.
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The present study demonstrates a unique mechanism for tumor cell-induced immunosuppression. In the presence of a nonsuppressive dose of tumor cells, generation of cytotoxic T cells in the mixed lymphocyte culture (MLC) is completely suppressed by adding exogenous (peritoneal) macrophages (PM phi) after the initiation of the MLC. This indicates that tumor cells can switch on a suppressor mechanism through host macrophages. It has further been determined that suppression can be induced only if resident (splenic) macrophages (SM phi) are exposed to tumor cells prior to addition of PM phi. If SM phi and PM phi are simultaneously present with the tumor cells, induction of suppression is completely precluded. These findings indicate that switching on of the suppressor mechanism by tumor cells has a critical requirement for the collaboration of two populations of macrophages, SM phi and PM phi, and their presence in a specific sequence (SM phi preceding PM phi). This may represent one of the mechanisms by which tumor cells evade host immune surveillance.
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Compared to normal hosts, tumor-bearing BALB/c mice were more susceptible to the immunosuppressive effect of tumor cells. At least a tenfold increase was found in the susceptibility mediated by a population of radioresistant spleen adherent cells (AC). The experiments were performed by the study of the suppressive effect of tumor cells on the generation of cytotoxic T-cells in allogeneic mixed lymphocyte culture reactions and allogeneic mixed lymphocyte tumor cell culture reactions. Fewer tumor cells were needed to suppress the T-cell-mediated cytotoxic responses of tumor bearers compared to normal hosts. A normal spleen population could be made to react like the tumor-bearing host by first depletion of its normal macrophages and then reconstitution with spleen AC from tumor bearers. Conversely, reconstitution of the macrophage-depleted tumor bearer's spleen with normal spleen AC made the tumor bearer react like the normal host. Furthermore, tumor cells were needed to trigger the spleen AC of the tumor bearer to fully exert their effect.
Depletion of macrophages from immune spleen cells by treatment with carbonyl iron and magnet or by in vivo treatment with carrageenan enhanced the in vitro secondary cell-mediated cytotoxic response against a syngeneic Friend virus-induced leukemia, FBL-3 cells of C57BL/6 mice. However, further depletion of macrophages by passing the carbonyl iron-treated immune spleen cells through a nylon wool column abrogated the cytotoxic response. The addition of splenic macrophage-enriched preparations from either FBL-3-immune or normal mice suppressed the cytotoxic response of immune spleen cells treated with carbonyl iron and magnet. This suppressive effect of splenic macrophages presented a marked contrast with the enhancing effect of normal peritoneal macrophages on the same cell-mediated cytotoxic response, indicating regulation of the generation of killer T cells against a syngeneic tumor by functionally distinct macrophages. The suppressed cell-mediated cytotoxic response against FBL-3 cells by immune spleen cells was augmented by the addition of indomethacin to the culture medium, and this augmentation with indomethacin was greatly decreased by depletion of phagocytic cells from the immune spleen by treatment with carbonyl iron and magnet. The mechanisms of regulation of the cell-mediated cytotoxic response with soluble factors released from macrophages are discussed.
High levels of primary proliferative response to a chemically induced sarcoma Meth A can be induced in syngeneic BALB/c spleen cells. Testing at the peak of proliferative response (2 days after sensitization in the mixed lymphocyte tumor cell culture), we found the responders to be resistant to anti-Thy 1.2 antibody lysis but susceptible to anti-Ia antibody lysis. When responders were subjected to various treatments before sensitization, it was found that removal of macrophages had no effect on the generation of proliferative response; high levels of proliferative response could be induced in enriched B cell preparations and in spleen cells from nude mice, but there was only a negligible amount of response in enriched T cell preparations. These findings indicate that the responders are primarily B lymphocytes. However, it was also found that the enriched B cell preparations usually gave only 50 to 75% of the response of whole spleen cells, whereas these B cells gave a 2- to 3-fold increase in the response to a B cell mitogen, LPS; this result indicate that collaboration from other types of lymphocytes was required for the generation of an optimal proliferative response to Meth A. Addition of 10% of T cells indeed produced a helper effect on this B cell response, and the maximal helper effect was seen for a mixture containing equal parts of T cells and B cells or for a slight T cell excess. These results indicate that the proliferative response to a syngeneic Meth A tumor is a macrophage-independent T-dependent B cell response.
In studying the immunogenicity of spleen cells and tumor cells in the generation, of cytotoxic T lymphocyte (CTL) in the allogeneic mixed lymphocyte culture (MLC) or mixed lymphocyte tumor cell culture (MLTC) reactions, we have found that the tumor cells not only appear to be poorly immunogenic, but are also immunosuppressive. This was shown by the ability of the tumor cells or their cell-free extracts to suppress standard MLC reactions. This suppression was acting mainly at the induction phase of the cytotoxic response. It could not interfere with the killing activity of the fully generated CTLs. In a Friend virus-induced leukemia FBL-3 system, at least two major components could be attributed to the cause of immunosuppression; one was of viral origin and the other was of non-viral origin. The viral component was sensitive to UV-irradiation and could be pelleted after ultracentrifugation at 100,000 g. The non-viral component was UV-resistant and was retained in the supernatant fraction after ultracentrifugation. Friend virus and 12 commonly found murine viruses have been excluded as the possible candidates causing the immunosuppression. The immunosuppressive viruses are very likely of endogenous origin and are defective in replication as shown by electromicroscopy, and by the virus focus-inducing and reverse transcriptase assays. These findings indicate that probably all tumor cells possess the immunosuppressive factor(s) which may account for their apparent lack of immunogenicity and the lack of proper immune responses in the tumor-bearing hosts.
The effect of macrophages on the induction of the cell-mediated cytotoxicity against a leukemia in a syngeneic system was investigated. The addition of exogenous peritoneal cells from normal C57BL/6 MIce enhanced the in vitro secondary cell-mediated cytotoxic response of both spleen and lymph node cells as responding cells against syngeneic FBL-3 leukemia. Peritoneal phagocytic macrophages seemed to be responsible for the enhancement. No inhibitory effect was demonstrated by the addition of peritoneal macrophages at a concentration as high as 20%, whereas the primary cytotoxic allograft response was significantly suppressed. In the present studies, there was no absolute restriction of macrophage-T cell interaction by an H-2 barrier. Supernatants of peritoneal macrophage cultures also enhanced this cell-mediated cytotoxic response. There was no difference between the effects of syngeneic or allogeneic peritoneal macrophage culture supernatants.
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By employing the 125IUdR release cytotoxicity assay, we have been able to measure the primary and secondary cell-mediated cytotoxic response of C57BL/6 mice to FBL-3 cells, a syngeneic Friend virus-induced leukemia. It was found that the secondary cell-mediated cytotoxic response occurred more rapidly after challenge (within 3 days) than the primary response, and the levels of reactivity were considerably higher. As in the primary response, the secondary cytotoxic reactivity of spleen cells was T cell dependent, being eliminated by pretreatment with anti-theta antibody plus complement. However, the secondary reactivity of pertioneal exudate (PE) cells was not entirely T-cell dependent. The specificity of the secondary cytotoxic response was analyzed by primary or secondary immunization with various tumor cells and by testing of cytotoxic lymphocytes against a variety of target cells. When spleen cells were used for testing, only tumor cells induced by Friend, Moloney, or Rauscher (FMR) leukemia viruses could produce secondary cell-mediated cytotoxic responses against FBL-3 cells. This correlated well with the specificity observed in the in vivo tumor transplantation protection studies. Similarly, spleen cells immune to FBL-3 had appreciable cytotoxicity against tumor cells induced by FMR viruses. The FBL-3 immune mice also gave significant protection against the challenge of FMR leukemias. When PE cells were used for testing, they gave higher levels of cytotoxicity against tumor cells induced by FMR viruses, but also gave less, but appreciable, cytotoxicity against non-FMR tumors. The latter reactivity might be related to the antigens induced by the murine endogenous type C viruses.
Cell-mediated cytotoxic reactivity of C57BL/6 mice against syngeneic FBL-3 cells, a Friend virus-induced leukemia, was measured by the 125IUdR release assay with tumor target cells in suspension. The tests could be performed either with Linbro tissue culture plates(16-mm wells) or with Microtest II plates (6-mm wells). The former could only be harvested manually; the latter could be harvested mechanically by an automatic harvesting apparatus which permitted larger scale tests. With either plate, it was found that careful preparation of the target cells and of the attacker cells has important effects on the results obtained. When performed under optimal test conditions, the 125IUdR assay can be used as a very simple, objective, and reproducible assay for testing cell-mediated cytotoxicity.
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OBJECTIVES: Describe cases of children with hepatosplenomegaly (HS) attended at the General Pediatric Teaching Ambulatory (AGER) of Instituto da Criança, São Paulo, identifying the main causes, evolution, necessity for hospitalization and/or referral to specialists. METHODOLOGY: Retrospective analysis of the records of children presenting HS on admission at AGER from September 1, 1993 to August 31, 1996. RESULTS: Of the 89 children included (age range, 1 to 148 months; 24 months), 64 (72%) were referred from other services for HS investigation. Most common presenting complaints were: fever--39 (44%); pallor--26 (29%); weight loss--21 (24%) and jaundice--14 (16%). Main alterations noticed on physical examination were: pallor--47 (53%) and short stature--17 (19%). Anemia was diagnosed in 70 children (79%); 35 children (39%) had infections; 7 (8%) metabolic disorders and 5 (6%) neoplastic disorders. The most frequent infections were of the urinary tract--9 (10%) and hepatitis A--6 (7%). Thirty six children (40%) were referred to specialists, 17 of which were already diagnosed. CONCLUSIONS: Most of the children with HS present deficiency anemia associated with infections which the general pediatrician is able to diagnose. Persistence of unexplained HS for more than 2 months, especially when there is substantial volume enlargement or alteration in the organs consistency, is an indication for referral to specialists.
The blue rubber nevus syndrome consists of multiple venous malformations in the skin and gastrointestinal tract associated with intestinal hemorrhage and iron deficiency anemia. Other organs may be involved. The causes of this syndrome are unknown. Its most common presentation is in the form of sporadic cases, but dominant autosomal inheritance has been described. It is a condition that affects both sexes equally, and its occurrence is rare in the black race. We present a case of this syndrome diagnosed in a 11-year-old patient. He had severe anemia and a venous swelling on the trunk. Similar lesions were found in the stomach, bowel, and on his foot. We emphasize the main clinical aspects: intestine, eyes, nasopharynx, parotids, lungs, liver, spleen, heart, brain, pleura, peritoneum, pericardium, skeletal muscles, bladder, and penis lesions, systemic complications that may occur to these patients which are thrombosis and calcification, as well as consumptive coagulopathy and thrombocytopenia that may occur within the nevi.
BACKGROUND: Breast hematomas are common after traumas, surgeries, or contusions. They are rarely spontaneous, but they can occur spontaneously in patients with hematologic disease or with coagulation disorders. MATERIAL AND METHODS: The authors report a clinical case of a 48-year-old female with a 27-year history of paroxysmal nocturnal hemoglobinuria who underwent mammography screening because of a painless palpable moveable node in the upper inner quadrant of the right breast. RESULTS: Mammography showed a partially defined heterogeneous node of 35 mm without microcalcifications in the upper inner quadrant of the right breast which, associated with the clinical features, seemed to be an hematoma. Further mammography and ultrasound after 45 days showed retrocession of the lesion, and another mammography obtained after 60 days was normal. Seventy-five days after the first episode, the patient complained of another node with a skin bruise in the upper outer quadrant of the same breast, which seemed to be a recurrent hematoma. Two months later the mammography obtained was normal. CONCLUSION: Breast hematoma must be thought of as a differential diagnosis for a breast node, regardless of previous trauma or hematologic disorders.
The Panará, who had previously lived in isolation from Brazilian national society in the Amazon forest, were first contacted in 1973. Two years later they were moved to another area in Central Brazil. During this same period they were reduced to 82 members, the survivors of a population of 400 to 500 in the mid-1960s. In 1995 they returned to a small area in their old territory still not occupied by outsiders. There, three years later, a health survey showed a presumed diagnosis of tuberculosis in 15 individuals out of a population of 181. Further tests in the town of Colider, based on clinical data and chest X-rays, confirmed the diagnosis in 10 Panará (6 children under 10 years of age and 4 adults from 40 to 50 years old). BCG scars were present in the entire population. The nutritional status of Panará children was better than that of other indigenous groups in the Amazon region. The following measures were introduced for Tb control: a) treatment follow-up in the village, under direct supervision by both a nurse and the local indigenous health worker; b) compliance with defined criteria for ending treatment; c) periodic control of contacts and non-contacts; c) and establishment of a reference system with the health services in Colider.