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Monoclonal antibodies (mAbs) are biomolecules of great scientific and practical significance. In contrast to polyclonal antibodies from immune sera, they are homogeneous and monospecific, since they are produced by hybridoma cells representing a clone arising from a single cell. The successful technology was described for the first time in 1975; the inventors were later awarded the Nobel Prize. Currently, mAbs are broadly used as a research tool, in diagnostics and medicine in particular for the treatment of cancer or in transplantology. About 47 therapeutics based on monoclonal antibodies are now available in the US and Europe, and the number is still growing. Production of monoclonal antibodies is a multistage, time-consuming and costly process. Growing demand for these molecules creates space for research focused on improvements in hybridoma technology. Lower costs, human labor, and time are important goals of these attempts. In this article, a brief review of current methods and their advances is given.

The paper reviews the known facts on the immunological response in infections with viral haemorrhagic fevers--dangerous pathogens for life and health of people. Immunological process registered in infections with viruses from Arenaviridae, Bunyaviridae, Filoviridae and Flaviviridae have been described. Moreover, the immunological response in infection with the RHD (rabbit haemorrhagic disease) virus from Caliciviridae family have been shown as a potential model for laboratory research on the duration and pathogenesis of viral haemorrhagic fevers.

In the study authors tried to estimate the diagnostic value ofAgNOR number and MIB-1 and PCNA expression in thyreocytes from various pathological lesions, with special attention to differential diagnosis of border-line lesions (nodule in adenomatus goiter--adenoma--follicular carcinoma). Fifty four thyroid glands were examined (10 adenomatous goiters, 20 follicular adenomas, 10 follicular carcinomas, 14 papillary carcinomas). Statistical analysis of obtained results allows in most cases to differentiate carcinomas from benign nodular lesions. The best differentiating factor for border-line lesions (especially differentiation between benign nodular lesions, neoplasms, nodules in adenomatous goiter adenomas and adenomas carcinomas) has turned out to be MIB-1. PCNA expression has differed significant between adenomatous goiter and follicular carcinoma and between adenoma and both carcinomas. AgNOR number has only helped in differentiation between nodules in adenomatous goiter and follicular carcinoma - correlations for other lesions have not been statistically significant.

Background and Purpose: All the types of the glial cells contain estrogen (ER) and progesterone receptors (PR) but their occurrence in glial tumors of the brain is still controversial. The aim of this research was the clinical analysis of ER and PR expression in correlation with histological malignancy and expression of p53 and PCNA., Material and Methods: The investigation was carried out on a group of 56 patients operated on at the Neurosurgical Department of Wroclaw Medical University. The percentage of tumors containing ER and PR was assessed and mean receptor expressions were compared. Classical histological tests, immunohistochemical tests for ER, PR, p53 and PCNA with monoclonal antibodies (DACO) were performed for every specimen of tumor tissue., Results: ER occurred in 24 cases (42.9%), PR in 10 cases (17.9%). 49% of highly malignant gliomas (WHO III and IV) were ER positive, whereas 29% of tumors grade I and II were ER positive. Frequencies of PR positive tumors were similar in both groups. Mean PR expression in p53 positive group was 8% and in p53 negative group 1.5% (p=0.017). Mean ER expression in PCNA positive group was 7.4%, whereas in PCNA negative group 2.7% (p<0.01)., Conclusions: Frequency of ER occurrence is higher in highly malignant tumors. ER expression is correlated with proliferative activity (PCNA). PR expression is positively correlated with intensity of mutant p53 protein.

Syndecan-1 is a member of the syndecan family of cell membrane heparan sulphate proteoglicans. The aim of this study was the evaluation of prognostic value of syndecan-1 expression in laryngeal cancer. The findings were correlated with the clinico-pathological parameters of the tumours, as well as with patient survival rate. Paraffin-embedded samples from 99 patients with laryngeal cancer selected from the files of the ENT-Dept. of Medical Academy in Lublin were immunostained with anti-syndecan-1 monoclonal antibody. The patients' mean age was 57 years and the over 5-year survival rate was 53.2%. Syndecan-1 immunoreactivity was observed in 99 (100%) of carcinomas. In our study, a statisti- cally significant correlation between syndecan-1 expression and patient survival rate was observed (chi2 = 9631; p = 0.008) as well as between syndecan-1 expression and various clinical stages of disease (chi2 = 6771; p = 0.034). The significant difference in the presence of syndecan-1 expression among the patients with various stage of histological differentiation of carcinoma (chi2 = 14.9; p = 0.001), and among the patients with present and absent metastatic changes in regional lymph nodes (chi2 = 16.698; p = 0.001) was observed. In a Coxs multivariate analysis syndecan-1 had an independent prognostic value (p = 0.014). Our results indicate that syndecan-1 could be used as a prognostic marker in laryngeal cancer.

Aberrant glycosylation is a universal feature of a cancer cell. The examples of such differences are the observed quantitative and qualitative changes in the expression of gangliosides of the tumors of the neuroectodermal origin. The role of gangliosides in cancer progression has been described, as well as their abilities to act as immunosuppressors. The presence of large amounts of these tumor associated carbohydrate antigens (TACA) on cancer cells, as compared to normal cells, opens the possibilities to use them in diagnosis and immunotherapeutic approaches which engage the immune system to fight with a tumor disease. The passive immunotherapy of neuroblastoma with anti-gangliosides monoclonal antibodies and their derivatives has been reviewed. The advantages and the disadvantages of using carbohydrate antigens as vaccines have been summarized. The examples of use of active specific immunotherapy with gangliosides have been described, as well as the approaches to modify the immunogenic potential of these antigens with carbohydrate-protein conjugate vaccines, and antiidiotypic antibodies used with immunomodulators such as QA-21. Finally, in a separate paragraph, the application of anti-carbohydrate antibodies to screen phage display peptide libraries for mimotopes has been described. The perspectives of using carbohydrate mimicking surrogate antigens in the immunotherapy of cancer have been discussed.

beta-catenin and APC are key components of Wnt signaling pathway. Moreover beta-catenin protein is an adhesion molecule, which exists in complex with E-cadherin. Although clinical impact of beta-catenin in laryngeal cancer is still unclear. Our purpose was to investigate an expression of beta-catenin and its possible prognostic value in laryngeal cancer. The group of 41 patients with laryngeal cancer, surgically treated with minimum 5 years observation, was multi-variously analysed. Paraffin-embedded tissue sections from each case were immunohistochemically stained with a monoclonal antibody raised against beta-catenin antigen. By univariate analysis expression of beta-catenin analysed on the front of laryngeal tumour proved to be significantly related to overall and disease free survival, which were shorter in cases with cytoplasmatic expression. Although there were co correlation with overall and disease free survival in assessment of beta-catenin examined in the middle of tumours. Expression of beta-catenin in the primary tumours was not associated with its size, nodal status, local and nodal recurrences and histological stage (grading). Analysis of beta-catenin protein expression enables the assessment of biology of laryngeal cancer and it can be a prognostic factor of overall and disease free survival, while measures on front tumour in patients with laryngeal cancer.

The aim of the study was to evaluate the clinical and prognostic significance of cytogenetic abnormalities in multiple myeloma patients. Cytogenetic studies were performed in 95 myeloma patients aged 31-82 (median 64) prior to chemotherapy. The GTG and CBG chromosome banding were performed and chromosomal abnormalities were described according to International System for Human Cytogenetic Nomenclature ISCN (1995). An abnormal karyotype was observed in 29% myeloma patients. Patients with an abnormal karyotypes showed various numerical and structural aberrations with hyperdiploidy in 39%, hypodiploidy in 39% and pseudodiploidy in 18% of patients. Monosomy of chromosome 13 was present in 29% of patients with an abnormal karyotype. Multiple myeloma patients with chromosomal abnormalities had more advanced disease than those with normal karyotype (82% vs 57% had stage III myeloma). beta 2-microglobulin and LDH levels were higher and hemoglobin level was significantly lower in patients with an abnormal karyotype. The plasma cell involvement of bone marrow was significantly higher in these patients. Overall survival was significantly shorter of patients with abnormal karyotypes (median 24 months vs 18 months), particularly of patients with monosomy of chromosome 13 (median 14 months). Cytogenetic studies are helpful to evaluate the prognosis and treatment options in multiple myeloma patients.

The percentages of CD4, CD8 subsets and Fas co-expression on them were assessed in secondary progressive multiple sclerosis (MS) patients and in control group. The MS patients were subdivided into two groups. The first group--active, consisted of patients who progressed by one or more degrees on EDSS scale in the preceding year, while the second group--stable, consisted of patients who showed no progression of the disease in the same period of time. Quantitative assessment of the individual subsets and Fas co-expression was carried out with the use of monoclonal anti-CD4, anti-CD8, as well as anti-CD95 antibodies, and Pas/Dako, Galaxy flow cytometer. There were no significant differences in CD4, CD8 subsets among the active, stable MS patients, and the control group. In the active MS group Fas co-expression on both subsets was significantly higher, as compared to controls. In the stable MS group, as compared to controls, a significantly higher Fas co-expression on CD8 subsets was observed. The investigation of Fas receptor expression on T lymphocytes in peripheral blood may be useful in order to monitor the progression of clinical activity in secondary progressive MS.

Background: Nephrotic Syndrome (NS) is a disease in which immune system function disorders play an important role. The aim of the study was an assessment of intracellular expression of cytokines in primary NS including assessment of intracellular expression of monokines (TNF-alpha, IL-8) in non-specific LPS stimulation system and intracellular expression of lymphokines (IL-2, INF-gamma, IL-4, IL-6) in experimental system with application of stimulation with ionomycin and PMA., Methods: The study included 47 children with NS aged 2-16 years. After activation of whole blood cells in the presence of brefeldin A the cells were labelled with monoclonal antibodies against surface antigens. In the second phase the cells underwent permeabilisation and were labelled with monoclonal antibodies against individual cytokines., Results: The studies performed demonstrated in children with NS relapse an increase of intracellular synthesis of cytokines characteristic of Th1 system (IL-2) and a decrease of synthesis of cytokines characteristic of Th2 system (IL-4, IL-6). In these patients an increase was also found of intracellular synthesis of proinflammatory monokines (TNF-alpha and IL-8). The studies performed, confirmed the earlier observations concerning abnormal cellular response in NS and demonstrated that differences in serum concentrations of individual cytokines may result from disturbances in their intracellular synthesis.

Unlabelled: The aim of the study was to characterize vascular stroma in glioblastoma (GB), to analyze its immunomorphological features, and to seek a relationship between patients' age and characteristics of the tumor microcirculation. Subjects in the study were 38 patients with GB, divided into two groups according to their age: group I--14 patients aged under 49 (from 32 to 49), and group II--24 patients over 50 years of age (51-78). Main types of vessels were specified: microvessels with normal morphology (capillaries, arterioles/venules), telangiectases, haemangioma--like forms, and microvascular proliferation types--simple and glomeruloid. An immunohistochemical analysis of the vascular stroma was performed with monoclonal antibodies against vWf and CD31, ASM, and Collagen IV (DAKO) to visualize the vessel wall components. The immunomorphological analysis was carried out for particular types of vessels separately. The vWf and ASM expression ratios--percentage of immunopositive area on 10 representative tumor fields under 100 x HPF--were assessed using a computer image analyzer. The incidence of some vessel types was dependent on the patients' age: in group I vascular--rich areas were more numerous, while in group II glomeruloid proliferation was more frequent--20/24-83.3% vs 8/14-57.1%. Immunoreactivity of the examined antigens was not related to the patients' age. The endothelial markers positivity was found within cells directly around the vascular lumen, and the reaction with vWf was more pronounced than CD31 reactivity. The most intense reactions were noted in proliferative changes of both types and in capillaries. ASM reactivity was found in abluminal cells within all types of vessels, most abundant in glomeruloid proliferation. ASM positivity was present also in some capillaries. Collagen IV staining revealed various abnormalities in the vascular basement membrane structure. No significant intergroup differences were found in the vWf and ASM expression ratios. However, in both groups the ASM expression ratio was significantly higher than that of vWf expression (p < 0.001)., Conclusion: Expression of the vascular wall antigens under study depends on the blood vessel type. The number of cells with smooth muscle phenotype is significantly higher than that of endothelial cells, especially within proliferative vascular changes. Microvascular changes intensity is higher in the older patient group.

The immunophenotyping of leukaemia and non-Hodgkin's lymphoma cells is based on staining the cells with monoclonal antibodies against surface and cytoplasmic determinants followed with flow cytometry analysis. The problems of immuno-phenotyping are associated with technical difficulties, changes in expression of determinants and the rare types of leukaemia and haematological disorders typical for newborns and infants. The lack of blast cells within cell suspension obtained for test may be the result of bone marrow disorder (aplastic anaemia, preleukaemic cytopenia) or technical pitfall. The changed expression of determinants on blastic cells observed as weak expression or overexpression or atypical combination of determinants requires a careful interpretation. In the diagnosis of rare types of acute leukaemia (e.g. erythroleukaemia, megakaryoblastic leukaemia, mixed lineage or undifferentiated leukaemia) the additional monoclonal antibodies beyond routine set are needed. A special concern is necessary in diagnosis of newborns and infants leukaemia or bone marrow disorders like myelodisplastic syndrome particularly in children with other systemic diseases e.g. congenital immunological deficiencies, Down's syndrome. The problems of immunophenotyping in non-Hodgkin's lymphoma are frequently associated with obtaining a representative material e.g. surgical tumour biopsy, lymph node. In some case the differential diagnosis including small round cell tumours and anaplastic type of lymphoma is necessary what requires an additional set of monoclonal antibodies. Despite of modern technology, morphology, immunophenotyping and histopathology remain the standard of complex diagnosis of lymphoproliferative diseases and haematopoietic disorders in children.

It is well known that classification of diabetes in the group of patients aged 25-55 years, particularly not obese, is a difficult clinical problem. The aim of the study was to estimate clinical status of the investigated group and evaluate biochemical features based on positive titers of humoral immunological markers in the development of diabetes mellitus. We studied 31 non obese, newly diagnosed patients (mean BMI < 26 kg/m2), mean age--41.6 +/- 11.1 years, mean duration of diabetes mellitus was 11.4 +/- 21.8 months. Nearly 50% of the patients aged 25-55 years showed presence of humoral immunological markers, which suggests high prevalence of autoimmunological diabetes. These patients have a lower reserve of endogen insulin secretion. The evaluation at both B-cell secretory capacity and markers of autoimmunity is recommended to provide a proper classification.

Unlabelled: The effective functions of human peripheral blood neutrophil are modulated by regulatory molecules depending on receptor agonists and adhesion molecules., Objectives: Evaluation of percentage of neutrophil adhesion molecules in pregnancy of diabetic mothers as well as their expression on peripheral blood neutrophils., Material: A-12 pregnant diabetic women at 7-13 and 34-36 weeks of first gestation, B-11 healthy pregnant women match for gestation, age and parity; C-11 healthy non-diabetic women; D-14 diabetic, non-pregnant women., Methods: MFI expression and percentage of CD18, CD11b, CD62L and CD54 were evaluated resting and activated by 30 min., fMLP preincubation using monoclonal antibodies anti-CD11b and anti-CD62L, employing Flow Cytometer FACSCalibur with computer program CELLquest., Results: Expression of CD11b and CD18 receptor on leukocyte of A, B, D groups was higher spontaneous and after stimulation by fMLP in I-st trimester compare to group C and then lowered in IIIrd. The significant difference in CD62L percentage was noticed resting and activated neutrophils in group A, which was lower compared to C and expression of CD62L on peripheral blood neutrophils was higher in group B compared to other studied groups in Ist trimester. There was an increase in expression of CD54 on resting neutrophils of group D when compared with C. A significant difference between Ist and IIIrd trimester was noticed in group A in expression of CD62L,CD62L/fMLP,CD54 on peripheral blood neutrophils, which were higher in the third trimester. Expression of CD62L was higher in the group A when compared to B., Conclusions: Diabetes type 1 and pregnancy changed the percentage and MFI of neutrophil adhesion molecules.

In 39 full-term neonates aged from 1 to 7 days of life, among them 24 with perinatal risk factors and 15 healthy as the control group, immunophenotyping of CD3+ and CD25+ cells using Becton Dickinson flow cytometer and monoclonal antibodies and CD3+ and CD25+ was performed. It was stated, that the mean relative and absolute number of CD3+ T lymphocytes in high risk neonates did not differ from the mean values in control group. The premature rupture of fetal membranes in mothers caused a significant increase of mean relative number of CD3+, whereas it did not influence on the absolute number of these cells. Significant increase of mean relative and absolute number of CD25+ cells in neonates with perinatal risk factors in comparison with the control group was noted. Mean relative and absolute number of CD25+ cells were significantly higher in neonates with non-infectious risk factors than in neonates with infectious risk factors. Authors emphasize that, evaluation of CD25+ cells is useful in the assessment of immunological changes in high risk neonates.

Unlabelled: In 27 infected neonates (16 boys and 11 girls) with birth weight ranged from 1200 to 4500 g (mean 2670 +/- 827 g) and gestational age from 31 to 41 weeks (mean 36.7 +/- 4.1), among them 11 (41%) with birth asphyxia, between 1 and 7 day of life flow cytometric immunophenotyping of the relative sizes of natural killer cells (CD3-/CD16-56+) using monoclonal antibodies of Becton Dickinson were made. The following infections were diagnosed in the neonates: bacterial septicemia (gram-positive in 13 cases, gram-negative in 5 cases), pneumonia in 6 cases and purulent meningitis in 3 cases. Control group consisted of full-term, healthy, eutrophic neonates born in Military Medical Academy in Lódź between 1994-95 years, in which immunological studies were made with the same method and dr Banasik, agreed for using her results for statistical analysis. It was stated, that in infected full-term neonates the mean relative size of NK cells was 10.7 +/- 5.1% and in prematures 12.8 +/- 7.2% and did'n differ significantly from the control. Kind of infection had no significant influence on the mean relative size of these cells, because in septic neonates was 11.7 +/- 5.7% and in other infected babies 11.6 +/- 7.2%. It was found that infected neonates without respiratory insufficiency and shock had significantly higher (13.2 +/- 6.4%) relative size of NK cells than healthy. The lowest value (7.0 +/- 4.8%) of NK cells during infections in dead neonates were noted (4 cases, among them 2 septic with shock and 2 pneumonic prematures with intraventricular haemorrhages)., Conclusion: The severity and bad results of the treatment in neonatal infections may be due to deficit of NK cells.

Objectives: EPH-gestosis is one of the most frequent complications of pregnancy, labour and puerperium. Despite its unknown ethiology, many authors suggest a vital role played by platelets as an ethiopathogenic factor. The aim of this study was to observe the level of platelets activation in EPH-gestosis subjects., Study Design: Platelets activation level has been observed in 16 EPH-gestosis third semester pregnant women and 14 normotensive third semester pregnant controls. Platelets double-labeled with monoclonal antibodies and a flow cytometer have been used in assessment of platelet activation level. Anti-P-selectin (GMP 140- Granule Membrane Protein) has been used as a marker of the platelet released reaction. Fibrinogen, D-dimers, antithrombin and thrombin-antithrombin complexes levels have been analyzed., Results: An increased platelet activation (p < 0.005) as well as the increased level of thrombin-antithrombin (p < 0.005) in pregnant EPH-gestosis women comparing to control group, has been observed in the study., Conclusions: A state of hypercoaguability and enhanced platelet activation in pregnant EPH-gestosis women have been observed.

It has been observed high risk of infections in neonates as a result from lymphocytes immaturity. It is connected to phenotype differences of lymphocytes between neonates and adults. This high susceptibility to infections is especially high in premature neonates. With the use of flow cytometry we have evaluated the phenotype of cord blood lymphocytes in premature neonates. In comparison to results of healthy newborns we have observed significant decrease in CD7+, CD3+, CD4+, CD25+, CD57+ lymphocytes an increase in total number of CD8+ cells as well as alteration in CD4/CD8 ratio. Our results suggest deeply damage of cellular immunity in preterm infants.

The authors analyzed the expression of oncogen and antioncogen proteins using immunoperoxidase technique with monoclonal antibodies in 49 primary ovarian carcinomas and 38 cancers after chemotherapy (second look) There was not found independent prognostic marker. But some results suggest, that in the individual cases estimate proliferative activity and overexpression P53 may give some information about intensitivity and quality of chemotherapy.

In this study we present evaluation of some immune parameters in children with borreliosis. We examined subsets of blood lymphocyte (CD3, CD4, CD8, CD19, CD3 HLA DR) and NK cells using monoclonal antibodies and flow cytometry in 14 children age from 3 to 16 years. We observed an altered lymphocyte phenotypes and NK cells in infected children compare to healthy control. We conclude that the changes in host immune response may play an important role in the pathogenesis of borreliosis and their monitoring may be helpful for the prognosis the course of the disease.

Myelin basic protein CD 2 lymphocyte stimulation index calculated as the proportion of percentage of myelin basic protein stimulated CD 2 lymphocytes to unstimulated CD 2 lymphocytes was evaluated in a group of 23 multiple sclerosis patients in the course of prednisone or methylprednisolone treatment. After both treatments a decrease of myelin basic protein CD 2 lymphocyte stimulation index was observed, statistically significant after methyloprednisolone medication.

Observation of percentages changes of lymphocytes (CD19), T(CD3), subsets CD4, CD8, NK cells, activated lymphocytes with fenotype CD3+HLA-DR, CD4/CD8 rate in Lyme borreliosis was carried out before (I examination) and after antibioticotherapy (II examination). 30 patients in aged 17-60 (x = 41) with recognition of erythema migrans(EM), Lyme arthritis(LA) and neuroborreliosis(NB) were examined. Epidemiological, clinical diagnosis was confirm by the presence of antibodies in ELISA test. Lymphocytes and their subsets, NK cells were signed and measured by flow cytometric immunophenotyping in Coulter EPIX XL with Becton-Dickinson antibodies twice: before and after treatment. Ampicillin, ceftriaxon or cefotaxim was applied during 4 weeks. AnStat program was used in statistic analysis. Value CD3 (x = 72.54) in I test was higher than control (x = 69.3), but CD19 (x = 13.2) was lower than control (x = 12.9). In II examination we stated CD19 (x = 9.48) progressive significant decreased in comparison to I examination. Percentage CD4 in II examination (x = 42) was lower than control (x = 45.8). Subset of CD8 had lower value in I examination (x = 28.03), as in II (x = 30.72) in comparison with control (x = 34.2). We showed lower CD4/CD8 (x = 1.40) rate after treatment than control (x = 2.67). We showed lower percentage NK cells after treatment (x = 13.5) than control. We also found lower percentage activated subsets T cells with phenotype CD3+HLA-DR before (x = 4.78) and after treatment (x = 4.03) compared with control (x = 7.27). No statistical changes in subsets with receptor for IL-2 before and after treatment was shown. In the course of active infection B.burgdorferi essential changes in lymphocytes subsets are observed. Decreased percentages of CD4, CD8, NK and CD3+HLA-DR+ in whole blood indicate their important role in immunopathogenesis of Lyme borreliosis. Lack of normalization of investigated parameters after treatment can be caused by inertion of elements immunologic system, as well as too short antibioticotherapies.

The aim of the study of 41 multiform glioblastomas was the analysis of p53-protein immunoreactivity in neoplastic cells and evaluation of relationship of this biologic marker to tumour proliferation activity. Positive p53 expression was observed in 24 (58.5%) tumours, the negative one in 17 tumours (41.5%). Proliferation indexes of PCNA, anti-Ki6 and AgNORs showed high values in the multiform glioblastoma p53 positive group, but without statistical differences in comparison with the group of p53-negative glioblastomas. Significant differences were observed in survival time of patients with p53 positive tumours in comparison with p53-negative ones. In 15 patients with p53-positive multiform glioblastomas survival time was less than 6 months (62.5%) on the contrary with only 4 patients with similar survival time in p53-negative glioblastoma group (23.5%). Our results suggest that p53 expression in multiform glioblastoma cells, generally considered as the indirect index of p53 suppressor gene, reflects aggressive stadium of neoplastic disease and significantly worsens the prognosis.

Sera of patients with acute and chronic hepatitis B and antigenemia were tested for the presence of anti-HBe and anti-HBs antibodies, free or bound in immune complexes. The possible occurrence of immune complexes of HBcAg in these sera was also investigated. Immune complexes were identified by antigen specific enzyme immunoassay, in which polyclonal antibodies against synthetic fragments of proteins S and C of HBV and mono- and polyclonal anti-HBc antibodies were used as a solid phase. Free and/or antigen bound anti-HBe antibodies were detected in 100% of patients with acute (81% HBeAg positive) and in 37% of patients with chronic hepatitis, all HBeAg positive. Anti-HBs antibodies or their immune complexes were found in 83% and 37% of patients, respectively. In not any patient circulating complexes of HBcAg could be identified. The results obtained support the observations that humoral immune response to HBeAg and HBsAg can be detected earlier than generally accepted; they also suggest that the detection of anti-HBs in a single sample of serum should not be considered as the evidence of elimination of infection.

Immunocytochemical analysis in fine-needle aspirates were performer in 46 cases of head and neck tumors. Immunocytochemical studies helped to establish the differential diagnosis of all diagnostically difficult cases. The application of 3 monoclonal antibodies against keratin, vimentin and LCA permitted to differentiate basic types of malignant tumors.

A case of medulloblastoma arising in advanced age is presented. Histologically it has anaplastic features despite desmoplastic character and astrocytic differentiation evidenced by glial fibrillary acidic protein immunoreactivity of some neoplastic cells. Neuroblastic differentiation could not be demonstrated by the neurofilament immunoreaction. A clear correlation between advanced age of the patient, the histological subtype of medulloblastoma and the direction of the tumor differentiation could not be established.

Expression of HLA-DR antigens by colon epithelium was examined in 35 patients with ulcerative colitis (uc) among whom 21 were in relapse and 14 in remission. Control group consisted of 18 subjects with functional disorders of alimentary tract. Immunohistochemical technique using immunoperoxidase was applied. Colon epithelial cells from all control subjects were HLA-DR negative. Cells of 17 out of 21 patients with ulcerative colitis in relapse and in 4 of 14 patients in remission were HLA-DR positive. No significant correlation was observed between the number of patients with HLA-DR positive epithelium and the insensitive of inflammation estimated clinically, endoscopically and histologically. 6 patients with initially HLA-DR positive epithelium during relapse were studied for the second time and in 4 of them the negative reaction was observed after remission occurred. The aberrant expression of HLA-DR may play an important role in the initiation and/or perpetuation of inflammatory process in ulcerative colitis.

IgG1, IgG2, IgG3 and IgG4 subclasses were detected in cerebrospinal fluid and sera from 32 patients with MS. The CSF levels of IgG1 and IgG3 subclasses in MS patients were significantly higher than those of controls. There were no statistical differences between the values of IgG subclasses in MS and control sera. The IgG1 and IgG3 indices were significantly higher in MS patients than in controls showing that among four IgG subclasses IgG1 ang IgG3 were synthesized in the central nervous system of patients with MS. The elevation of IgG1 and IgG3 indices in MS patients was found more frequently (in about 90% of patients) than the elevation of the general IgG index (in 72% of patients).

Platelet specific antigens and platelet antigens shared with other peripheral blood cells are presented. The molecular characteristics of membrane glycoproteins which are platelet specific determinants is discussed.

Surface expression of class II antigens (HLA-D) on B lymphocytes was studied in 47 patients with chronic lymphatic leukemia (CLL) with an aid of monoclonal antibodies against monomorphic determinants of DR, DP and DQ. An association was found between progression of disease and gradual loss of those markers, especially DP and DQ.

Paper presents the progress in antigenic characterization of different histological types of ovarian carcinomas. The role of monoclonal antibodies is pointed out. Clinical significance of tumor-associated antigens and problems of tumor cell heterogeneity for immunological studies are also reviewed.

102 Chlamydia trachomatis isolates obtained from pregnant women and 42 isolates from man-consorts of women with chlamydial infection of cervix were immunotyped using a microimmunofluorescence (micro IF) with monoclonal antibodies kit (Washington Research Foundation, Seattle, USA). In both groups of patients the most common serovars were: E (37.3%) and D (24.6%) belonging to B-complex. Additional serovars noted were: F (11.3%), J (11.3%), I' (4.5%), I (3.5%, K (3.5%), G (2.8%) and H. Ba (both 0.2%). N. gonorrhoeae and/or T. vaginalis infections were more frequent from patients with B-complex Ch. trachomatis serovars (28/75 that is 37%) than C-complex Ch. trachomatis serovars (3/25 that is 12%).

Immunochemical investigations of ribosomes should correlate with basic knowledge of the function, structure and activity of organelles in the cell processes. Our paper presents data of immunochemical methods used to determine the structure, function and differences of ribosomes. We present the usefulness of immunochemical methods to test human ribosomes, diagnosis and therapy of many diseases.

In this study, hepatic microsomes from 5,6-benzoflavone induced C57BL/10 mice were used. To inhibit monooxygenase activities, the monoclonal antibody MAb 1-7-1 recognizing two isoenzymes of methylcholanthrene-induced cytochrome P-450 was applied. Microsomes were incubated with tritium labeled benzo(a)pyrene [G-3H]BP for 10 min at 37 degrees C. The incubation mixture contained: 50 mM potassium phosphate buffer, pH 7.25; 30 mM KCl; 3 mM MgCl2; 2 mM NADPH; 80 microM [G-3H]BP (specific activity 50 mCi/mmol); and monoclonal antibody MAb 1-7-1 or ascites fluid (NBS) containing nonspecific IgG as a control. The ratio of antibody protein/microsomal protein was 2:5. BP metabolites were extracted from incubation mixtures by ethyl acetate. The organic layer was dried over sodium sulfate, and evaporated under a stream of nitrogen. To separate BP metabolites HPLC technology was used. The column was eluted with methanol gradient (60-100%) for 45 minutes. The radio-activity of collected samples was determined using liquid scintillation counter. Differential inhibitory effects of MAb 1-7-1 on BP-metabolites formation were found, e.g. 7,8-diol was inhibited by 86.1% and quinones by 62.5%. The predominant metabolite, 3-OH-BP, was inhibited by 80.4%. Moreover, it was found that MAb 1-7-1 inhibition of benzo(a)pyrene hydroxylase activity (by 75.8%, as measured by fluorescent technique) was very similar to the inhibition of 3-OH-BP along with 9-OH-BP formation (as measured by HPLC).

A case is presented of a rare T-phenotype acute lymphoblastic leukaemia. The blast cells were negative in the rosette test and demonstrated expression of the Ia antigen.

Monoclonal antibody 791T/36 directed against surface antigen on the human osteosarcoma cell line and cross-reacting with surface antigen p72 presenting on human PHA-stimulated T-lymphoblasts was used in analysis of p72 antigen expression on human mononuclear peripheral blood cells, between 2-4 days of culture. Using FACS-IV system and fluorescence microscope, it was shown that expression of p72-antigen is dependent on PHA-stimulation and the ability of lymphocytes proliferation. The expression of p72 preceding the entry of the cells into the cell cycle. This is not significantly dependent of cell size, stage of the cycle, and RNA-transcription activity. In PHA-stimulated cultures, between 2-4 days, the number of lymphoblasts expressing enough receptors for 791T/36 monoclonal antibody is sufficient to exhibit a distinct effect of Ricin A-chain conjugated with 791T/36 over 50% inhibition of 3H-Thymidine incorporation into the cells. These observations emphasises the importance of cross-reactions in cases using immunotoxins.

This paper presents a review of studies on the structure and function of receptors for biologically active substances (hormones, drugs etc.). Application of the methods utilizing monoclonal antibodies are described and discussed.