[Hypnosis for rehabilitation of immunological status in neoplasia].

The study group included 21 patients with malignant melanoma stage II-IV, aged 25-67, and 25 patients, aged 28-68, (control) with stomach tumors stage I-IV. All patients received individually-tailored hypnosis. Our newly-developed methods used batteries of suggestive images to deal with non-psychotic disorders and to map out strategies to support immunocompromised patients. In group 1, suggestion stimulated the "devouring" effect of the "patroling" cells. Quantitative and qualitative characteristics of immunocompetent cells of peripheral blood were assessed by flow cytometry, immuno-enzymatic analysis and other procedures to evaluate immunological status. Correlation analysis of data on group 1 identified 7 negative coefficients (p < 0.05 and p < 0.01) exactly in the monocytic macrophageal link thus suggesting the modulating effect of hypnosis. Our results support evidence available on the potential of hypnosis for cancer patient immunity and point for the first time to feasibility of differentiated targeting specific links of the immune system.

Characterization of immunogenic properties of polyclonal T cell vaccine intended for the treatment of rheumatoid arthritis.

Two-staged technology for obtaining polyclonal T cell vaccine intended for the treatment of rheumatoid arthritis is described. Stage 1 includes antigen-dependent cultural selection of patient's T cells and stage 2 consists in their reproduction in the needed amounts by nonspecific mitogenic stimulation. T cell vaccination induces an effective specific anti-idiotypic immune response against T cells reactive to joint antigens. Vaccine therapy significantly reduces plasma level of IFN-gamma and increases IL-4 level. The results indicate immunological efficiency and safety of polyclonal T cell vaccine in patients with rheumatoid arthritis.

Cell therapy of comatose states.

We demonstrated that liquor from adult humans can maintain proliferative activity of cells of immature nervous tissue in vitro. The paper presents the results of a retrospective clinical study of the efficiency of cell therapy in the treatment of II-III degree comatose patients with severe brain injury. Cell suspension consisting of cells derived from immature nervous and hemopoietic tissues was injected into the recipient subarachnoidal space through a cerebrospinal puncture. The mortality in the study group was 8% vs. 56% in the control group. The 1.5-year follow-up demonstrated significantly better quality of life in patients receiving cell therapy in comparison with patients of the control group. Cell therapy proved to be ineffective for patients in a comatose state caused by hypoxic encephalopathy. The study demonstrated the efficiency of cell therapy in patients with severe brain injury during the acute period of the disease.

Cell therapy of brain stroke.

Cell suspension consisting of cells from immature nervous and hemopoietic tissues was subarachnoidally transplanted to 10 patients with brain stroke consequences. Clinical effect of different degree was attained in all patients. Six months after cell therapy functional activity significantly increased in contrast to clinically comparable control group. No serious complications of cell therapy were observed. Presumably, cell therapy is a more or less safe method of treatment, which can be effectively used in the treatment of brain stroke consequences.

Erythroid cells in suppressing leukemia cell growth.

This paper indicates that murine nucleated erythroid cells (EC) are able to reduce, in a dose-dependent manner, the proliferation of both L1210 lymphoma and P815 mastocytoma cells and that the leukemia cell growth inhibitory activity of unseparated bone marrow (BM) cells may be markedly augmented by their short-term culturing with erythropoietin (Epo). These results raise the intriguing possibility to utilize erythropoesis-stimulating, therapeutic strategies with the purpose of inhibiting leukemia cell growth in the body.

Cell therapy of cerebral palsy.

The paper presents the results of a controlled study of cell therapy in 30 patients with severe forms of cerebral palsy. A cell suspension from immature nervous and hemopoietic tissues was injected into the subarachnoidal space of a recipient through a spinal puncture. Immune sensitization to donor antigens (detected by suppression of lymphocyte migration) was noted in few patients. In none patients laboratory and clinical signs of tissue-destructive autoimmune reactions were observed. One year after treatment activity of the major psychomotor functions in treated patients considerably surpassed the normal. No delayed complications of cell therapy were noted. These findings suggest that cell therapy is an effective, safe, and immunologically justified method of therapy for patients with cerebral palsy.

Characterization of erythroid cell-derived natural suppressor activity.

Nucleated erythroid cells (NEC) have been previously reported to the capable of suppressing antibody-mediated primary (IgM) and secondary (IgG) immune responses to thymus-dependent antigens. In the present study we indicated that NEC, separated from the spleens of mice following phenylhydrazine treatment were able to suppress directly the proliferative response of preactivated B cells to lipopolysaccharide (LPS) in vitro. While being active in suppressing B cell blastogenesis, NEC, however, failed to reduce both cell proliferation and cytotoxic T lymphocyte (CTL) generation in an allogeneic mixed lymphocyte culture (MLC). NEC also lacked a significant effect on interleukin (IL)-2 production and utilization by concanavalin A (Con A)-activated T lymphocytes. The NEC-derived suppression of B cell proliferation was, at least in part, mediated by soluble molecules. The specific blockade of transforming growth factor (TGF)-beta synthesis with antisense oligodeoxynucleotides (OD) binding TGF-beta mRNA, as well as the neutralization of TGF-beta activity with anti-TGF-beta antibodies (Ab), resulted in a detectable diminished ability of the NEC-conditioned medium (CM) to suppress B cell blastogenesis. Taken together, the results suggest that: 1) NEC may suppress directly B cell responses, while not affecting T cell ones; 2) NEC may mediate their natural suppressor (NS) activity partially through releasing TGF-beta.

Induction of mixed allogeneic chimerism for leukemia.

Hybrid (C56BL/6 x DBA) (BDF1; H-2b/H-2d) mice bearing the P815 leukemia (H-2d) were grafted with a (CBA x C57BL/6)F1 (CBF1; H-2k/H-2b) cell suspension, comprising bone marrow cells (BMC; 25 x 10(6)/mouse) and spleen cells (SC; 55 x 10(6)/mouse) on day-4, then treated with cyclophosphamide (200 mg/kg) on day-2 and finally grafted once more with CBF1 cells (25 x 10(6) BMC + 7 x 10(6) SC) on day 0. Allogeneic cell graftings performed in this way induced durable mixed hematopoietic chimerism and significantly prolonged the survival of recipients, compared with that of leukemia-bearers grafted with syngeneic cells. The results obtained raise the possibility of using allogeneic hematopoietic tissue transplantation in combination with non-lethal cytoreductive therapy to induce a long-lasting graft-vs-leukemia effect.

A role for interferon-gamma and transforming growth factor-beta in erythroid cell-mediated regulation of nitric oxide production in macrophages.

Interferon-gamma (IFN-gamma) and transforming growth factor-beta (TGF-beta) are known to be a potent inducer and inhibitor for macrophage (Mo) activation process, respectively. In the present study we established that the nucleated erythroid cells (NEC) separated from the spleens of adult (CBA x C57BL/6)F1 (CBF, H-2k/H-2d) mice following phenylhydrazine treatment are potentially capable of inducing nitric oxide (NO) production in thioglycollate broth-elicited peritoneal macrophages (Mo). The stimulating effect of both NEC and their culture supernatant on NO secretion by Mo was most apparent in the presence of bacterial lipopolysaccharide (LPS) and neutralizing antibodies (Abs) to TGF-beta and was largely reversed by the addition to the culture of neutralizing Abs to IFN-gamma. Collectively these results suggest that NEC, through production of IFN-gamma and TGF-beta, may exert a regulatory influence on development and functionality of cells pertaining to monocyte (Mc)/Mo lineage.

Tumor growth inhibitory and natural suppressor activities of murine bone marrow cells: a comparative study.

The murine bone marrow (BM) cells having a certain phenotypic similarity to null natural suppressor (NS) cells have been previously established to be able to inhibit in vitro leukemic cell growth in a genetically unrestricted manner. In this study we found that the treatment of normal (C57BL/6 x DBA)F1 BM cells with a lysosomotropic agent, L-leucine methyl ester (LME), largely abrogated their ability to reduce both P815 mastocytoma and L1210 lymphoma cell proliferation, as well as their NS activity tested for suppression of mitogen (Con A or LPS)-driven spleen cell proliferation. However, after being depleted of the cells binding wheat germ agglutinin (WGA), the BM cells maintained tumor growth-inhibitory activity, while demonstrating no significant NS activity. Moreover, in contrast to T-cell blastogenesis-inhibitory NS activity of BM cells, that was greatly reduced by the addition into the culture of either neutralizing anti-interferon (IFN)-gamma antibody (Ab) or NG-monomethyl-L-arginine, a competitive inhibitor of NO synthase, natural antitumor cytostatic activity of BM cells was not found to be dependent on the presence in medium of IFN-gamma and to be associated with NO production. When incubated at suboptimal numbers with tumor cells on conic, round, and flat well bottoms for 7 h, BM cells provided the most, middle, and least (or no) tumor growth inhibition, respectively, suggesting, thereby, a significance of cell density in cytostatic process. It was also found that the BM cells cultured for 20 h with the medium conditioned by mitogen-preactivated T or B lymphocytes were significantly more suppressive to tumor cell proliferation than the BM cells cultured in medium alone. The potentiation of BM-cell cytostatic activity by T-cell-conditioned medium (CM), but not that by B-cell-CM, was found to be partially reversed by anti-IFN-gamma Ab. Finally, a noticeable tumor growth-inhibitory activity, which could be significantly enhanced upon T-cell-CM, was shown to be also attributable to BM cells from athymic BALB/c mice. Taken together, the results suggest that (1) the tumor growth inhibitory BM cells and the NS BM cells are not identical in their cell compositions, but also differ in their mechanisms of antiproliferative action; (2) a contact cell-to-cell interaction may play a significant role in BM-cell-mediated tumor growth inhibition; (3) the activated lymphocytes, through both IFN-gamma-mediated and IFN-gamma-independent pathways, are able to operatively up-regulate the cytostatic activity of BM cells; and (4) the tumor growth-inhibitory activity exhibited by the normal unmanipulated BM cells, at least in its significant part, may not be a consequence of thymus-dependent immune processes occurring normally in the body.

Cytokine gene expression in erythroid cells.

Erythroid nuclear cells have been shown to exert regulatory effects on immunopoiesis. We have reported that some of these influences might be mediated via soluble factors secreted by nuclear erythroid cells. In this report we describe our estimate of the cytokine gene expression in cells isolated from individual erythroid colonies by Reverse transcription-Polymerase chain reaction. We found in erythroid cells, originated from the bone marrow precursors obtained from phenylhydrazine-treated mouse, the expression of the following cytokine genes: IL-1 alpha and IL-1 beta, IL-4, IL-6, GM-CSF, gamma-IFN and TGF-beta. In contrast, the erythroid cells derived from newborn mouse spleen precursor cells expressed IL-1 alpha, IL-1 beta, IL-4, IL-6 and GM-CSF mRNAs but not gamma-IFN and TGF-beta mRNAs. No detectable levels of IL-2, IL-3 and IL-5 mRNAs were expressed in nuclear erythroid cells. These data provide evidence of the expression of mRNAs coding in the set of immunoregulatory cytokines in immature erythroid progenitor cells in mouse.

Bone marrow cells as cytostatic effectors responsible for suppressing leukemia growth in vitro.

When bone marrow (BM) cells, isolated from normal (C57BL/6 x DBA/2)F1 mice (H-2b/H-2d), were cultured with leukemic cells for 24 hours, a significant tumor growth suppression, without noticeable tumor cell killing, was found. The level of BM cell-mediated cytostasis of both P815 mastocytoma (H-2d) and L1210 lymphoma (H-2d) cells was dependent on BM-to-tumor cell ratio; 100% growth inhibition was obtained at a ratio of 480/1. In addition, BM cells were found to be able to synergize in suppressing P815 cell growth with lymphoid cells. The synergistic suppressive effects on tumor cell proliferation were observed in BM-spleen, BM-thymus and BM-lymphnode cell co-cultures. The analysis of cytostatic activity of the cell culture supernatants showed that the synergistic leukemia growth suppression could be mediated, at least in part, by cell-derived soluble cytostatic molecules. The data presented herein also indicated that culturing BM cells with either crude supernatant (25%) from allogeneic mixed lymphocyte culture (MLC) or recombinant human interleukin(IL)-2 (20 U/ml) for 20 hours led to a 2-fold increase in their cytostatic activity against both P815 and L1210 cells. Taken together, the results suggest that although normal BM cells are ineffective in tumor cell killing, they may play an important role in cell-mediated effector mechanisms responsible for suppressing leukemia development; and that activated T lymphocytes, through producing cytokine(s), may rapidly upregulate leukemia growth inhibitory activity of BM cells.