Search

Osteosarcoma (OS) is the most frequent primary malignant bone tumour, mainly affecting children in the first and second decade of life. Causes of the disease are still unknown and reaction of the immune system on its development is very individual. Particular emphasis must be placed on the role of cytokines in the immunoregulatory and coordinating function and tumour cell disruption. Knowledge about cytokines concentration in serum, regarding mechanisms of oncogenesis, may have prognostic significance for the further course of OS in children. The aim of study was evaluation of IL-2, IL-4, IL-8, IFN-gamma, and TNF-a concentrations in children with osteosarcoma at diagnosis.The study was performed on the group of44 children with osteosarcoma, aged from 6 to 20 years (average 14.9 years; median 15.0 years). 22 children ofthesame age (median 14.5years) without neoplastic disease and active inflammatory state formed the control group. Investigations were performed before the therapy. The inclusion criteria were: diagnosis of primary osteosarcoma, upper or lower limbs tumour localization, patients who were not treated by chemo- or radiotherapy before biopsy patients' age at diagnosis was 6-20 years. Concentrations of selected cytokines were analyzed in peripheral blood with using ELISA method with 99.8% sensitivity and 99,5% specificity.In children with osteosarcoma, at diagnosis the following concentration of peripheral blood cytokines (medians) was observed: IL-2 10.7 pg/ml (min-max: 0.0-894.0); IFN-gamma 1,3 pg/ml (min-max: 0.2-147); TNF-alpha 28.3 pg/ml (min-max: 0.0-188.8); IL-4 2.0 pg/ml (min-max: 0.0-32.0); IL-8 13.5 pg/ml (min-max: 0.0-2154.0). A large scatter among individual children results was found. Analysis of cytokines concentration showed significant statistical differences between patients with OS and the control group in case of IL-4 (p=0.005) and IL-8 (p=0.01).Results of studies obtained at diagnosis did not give a specific answer about the prognosis and further course of OS disease in patients in the developmental age. Big differences in cytokines concentration in children and youth with OS might be associated with individual biological variation and individual reaction to the development of neoplastic disease and further studies in this direction are needed - before the start of cytostatic therapy and in therapy monitoring.

Food allergy is an abnormal response of the immunological system, especially of mucosal immunological system on antigens supplied per os. There are very complicated and still unexplained immunological mechanisms, which lead to hypersensitivity reaction. Most often food hypersensitivity is identified as the effect of atopy, which is connected with humoral response (specific IgE antibody). On the other hand cell immunological response are less investigated, however they can be very important, especially as a significant factor to initiate pathological allergic processes.To investigate, the usefulness of flow cytometry for estimation of specific sensitization of subpopulation of lymphocytes on food allergens in the allergy diagnosis.The investigations were performed on 60 children from 6 months to 5 years old: 20 children with CM A IgE dependent, 20 with CM A IgE independent and 20 healthy children. IgE total, sIgE, IgG, IgA, IgM, basic immunological panel, CD 23, CD25, CD26, CD30, CD69, PCNA were measured.We noticed decrease of expression of CD4+CD30+ between I and II examination (p=0.029), between I and III (p=0.009); decrease of expression of CD8+CD26+ between I and III test (p=0.038); decrease of expression of CD19+CD23+ between I and II examination (p=0.012) in I type of hypersensitivity. We observed a decrease of expression of CD4+CD25+ between I and III examine (p=0.026) and decrease of expression of CD4+ CD26+ between I and III examination (p=0.036) in IV type of hypersensitivity. Expression of CD69 was decreased after diet in IgE dependent allergy. Values of expression of PCNA are similar in I and IV type of hypersensitivity in children with CM A. Decrease of expression of PCNA in II examination was observed in both cases. Reintroduced allergen caused increase of expression of PCNA in both types of allergy (p=0.048 and p=0.041).Our recent research confirms changes of the expression of T lymphocytes activation markers. It is connected with in vivo stimulation to allergen or with allergen elimination. The study of expression of activation markers using flow cytometry in food allergy in children can be helpful in observation of the dynamic progress process, but it cannot be used as a single diagnosis test.

Many studies in vitro and in vivo have shown immunomodulating and antiviral activities of inosine pranobex. The object of this research was to examine the potential beneficial effects of inosine pranobex (Groprinosin) on immune system in children with cellular immunodeficiency as a prophylaxis of recurrent infections, mainly of viral origin. 50 mg/kg b.w/day of inosine pranobex in divided doses was given to the group of 30 children aged 3-15 years for 10 days in 3 following months. Clinical and immunological investigations were done before and after the treatment. Statistically significant rise of CD3T lymphocytes number (p = 0.02) and in this CD4T lymphocytes number (p = 0.02) as well as statistically significant improvement of their function (p = 0.005) evaluated with blastic transformation method were found. These laboratory findings were parallel to clinical benefits. Control study was performed in the group of children completed by randomization and treated in the same way with garlic (Alliofil).

The immonomodulatory mechanisms responsible for allergens tolerance in healthy subjects were described on the basis of the newest medical literature, including own papers and experience. It was detected that the same immunological mechanisms are built by specific immunotherapy. IL-10 and TGF-beta cooperate in the regulatory T-cell response to aeroallergens, food allergens and allergens of bee and yellow jacket venoms in healthy subjects as well as during specific immunotherapy making the tolerance to these allergens. The actual progress in improvement of specific immunotherapy methods is described.

As the frequency of allergic disease and especially the food allergy in children is still increasing - we undertook studies to determine the specific sensitivity of lymphocytes T in order to improve the diagnostics and monitoring of treatment. The usefulness of evaluation CD69 and PCNA expression of T cells from children with cow's milk allergy (CMA) in these procedures was examined.The study on usefulness of lymphocytes CD69 and PCNA detection for diagnosis of CMA in children and for monitoring of the treatment.The subjects in this study included 26 children from 1.5 to 15 years old with CMA who were under clinical care of our Immunological Department. Patients were examined (medical history of the child, physical examination, specific food provocation) and laboratory investigations were made before therapy and after 6 months of elimination diet. Allergen-specific IgE antibody was determined. Children's T cell cultures were stimulated by beta-lactoglobulin, alpha-casein, whole cow's milk, phytohaemoglutinin PHA and without stimulation. Expression of T cells CD69 and PCNA from patients with CMA and 10 healthy children were analysed by use of flow cytometry. 10 children with CMA after 6 months of elimination diet and next 2 weeks with cow's milk provocation were examined also for CD69 and PCNA expression in lymphocytes from peripheral blood.After 6 months of elimination diet it seems that the expression of CD69 receptor has tendency to fall, these differences, however, are not statistically significant. But after next 2 weeks with cow's milk diet the expression of CD69 was increased with statistically significance (p=0.04). T cells PCNA expression of children with CMA was lower than in healthy children in PHA stimulated cultures. Our study indicates that PCNA expression of T cells from patients with CMA increased even more after 6 months of elimination diet in all cultures but there was statistically significant differences only after stimulation with cow's milk and PHA (p=0.05 and p=0.04 - respectively). Subsequently through 2 weeks children were provoked with cow's milk and after that PCNA expression in vivo lowered (no statistically significant differences).CD69 expression of T lymphocytes was significantly higher after specific stimulation of children with cow's milk allergy (p=0.04). The evaluation of CD69 expression of children with CMA may be also useful in monitoring of treatment. Estimation of PCNA expression suggests the efficiency of T cells connected with DNA synthesis related to the stage of allergy disease and demands further investigations.

The possibility of detection of lymphocytes T specific sensitivity after contact with the allergen (mainly food allergen) even in early age of children has been reported in this paper.This is the literature review of basic applied methods such as: lymphocyte blastic transformation in allergen stimulated culture, macrophage migration inhibition test and flow cytometric analysis of in vivo and in vitro lymphocyte specific activation by using activation markers (CD69, PCNA - proliferating cell nuclear antigen and others).The significance of such investigation is underlined especially for young children during early diagnosis of food allergy and monitoring of the treatment.