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AIM: A regular monitoring of the immune reconstitution mainly based on the quantitative determination of lymphocyte T subpopulation. This is prospective analysis for 1 year in Tunisian children treated with allogenic intrafamilial bone marrow transplantation. Methods: We conducted a prospective analysis for 1 year follow up enrolling 25 children treated with allogenic intrafamilial bone marrow transplantation among them two cases of Peripheral hematopoietic transplantation and placental cord blood transplantation including: aplastic anemia (6 cases), hemoglobinopathies (12 cases), myelodysplastic syndrome (1 case), 2 cases of Acute lymphocytic leukemia, a case of congenital amegacarycytosis and 3 cases of primary immunodeficiency with lack of expression of major MHC class II. All subjects received different conditioning regimens according to the indication. Our study consisted of a regular monitoring of the immune reconstitution mainly based on the quantitative determination of lymphocyte T subpopulation. So, these tests were routinely requested to 1 month, 2 months, 3 months, 6 months, 9 months and 12 months post- bone marrow transplantation. Results: The average time of engraftment was 18 days corresponding to neutrophil recovery (12-24). For the T cell recovery, a rate of CD4 + T lymphocytes > 200/ mm3 was provided within an average of 2.5 months (1-7). The average time to obtain CD8+ T lymphocytes >200 /mm3 was 2 months (1-5). The humoral immune reconstitution was made within an average of 2 months (1-4). A ratio of CD4+ / CD8+ T lymphocytes (>1) was obtained within 10 months and a half (1-24). Univaried analysis showed a significant correlation between the bone marrow sex matched and the faster reorganization of CD8 + T cells (p = 0.042). Moreover, a quantity of CD34 +> 6x 106/ kg was significantly associated with the recapture of a formula lymphocyte T CD4+ / CD8+ (> 1) (p=0.03). Conclusion: The immune recovery post bone marrow transplantation in children began with myeloid lineage then lymphoid B then lymphoid T. The inversion of the ratio CD4 +/CD8+ T lymphocytes, seemed to be influenced on the one hand by the high content of CD34 + cells in the graft as well as the type of conditioning on the other hand by the CMV infection since it accelerates significantly CD8+ T lymphocyte reconstitution.

This study compared retrospectively the effectiveness, toxicity and hematopoietic recovery after autologous peripheral blood stem cell transplantation (ASCT) of two consecutive peripheral blood stem cell mobilization regimens in newly diagnosed MM patients. Patients in group 1 (n=178) were treated with 4 g/m

We explored the influence of polymorphisms in genes encoding the chemokine stromal cell–derived factor-1 (SDF-1)/CXCL12 in a cohort of Tunisian patients with malignant hematologic diseases multiple myeloma [MM], non-Hodgkin's lymphoma [NHL], Hodgkin's disease, and acute myeloid leukemia [AML], who underwent stem cell mobilization for autologous transplantation versus a group of healthy donors for allogeneic transplantation. Polymerase chain reaction–restriction fragment length polymorphism (PCR-RFLp) analysis was used for rapid identification of genotypes. Significant associations for SDF1-3′A polymorphism were observed exclusively in patients with MM and NHL. While there was a lack of all association of SDF-1 polymorphism with AML patients. However, considering that the ability of mobilization varies among subjects, we have observed that the SDF1-3′A allele was associated with good mobilization capacity. Interestingly, the association was mainly observed among healthy allogeneic transplant donors where the analysis was not biased by background disease or chemotherapy (P = .010; odds ratio = 2.603; confidence interval [95%] = 1.239–5.466).

Apoptosis represents a particular form of programmed cell death which appears in all the damaged cells and potentially hazardous. It plays a crucial role in the development of multicellular organisms by assuring and maintaining the cellular homeostasis. Thus, apoptosis intervenes not only in the normal process of organisms' development but also in immune defence and in cancerous cells detection. Indeed, any blockage in the program of the apoptotic machinery would be responsible of some neurodegenerative and auto-immune diseases and could play a crucial role in different steps of carcinogenesis. Some researchers were very interested in studying apoptosis in hematopoietic stem cells CD34+ which could be intended to be reinfused to patients suffering from malignant diseases. They have noted that kinetic study of apoptosis of the hematopoietic stem cells CD34+ after the process of cryoconservation is also necessary. Such study permits to quantify the real and exact number of the viable hematopoietic stem cells CD34+ and therefore to eliminate such risk which would be associated with the reinfusion of apoptotic cells to patients. In this paper, we describe our contribution to hematopoietic stem cells CD34+ study by flow cytometry before and after cryopreservation by using annexin V as a specific probe allowing detection of phosphatidyl serine, one of the major features of apoptosis. But, we have noted a pronounced induction of apoptosis in peripheral mobilized blood compared to cytapheresis (after cryopreservation: 29.79% of apoptotic HSC CD34+ in peripheral mobilized blood but only 11.67% apoptotic HSC CD34+ in cytapheresis). Besides, we have noticed that hematopoietic stem cells CD34+ have had a statute of viability better than other mononuclear cells. These results put in value the reliability, the simplicity and the efficiency of flow cytometry for the analysis of apoptosis in hematopoietic stem cells CD34+ by following the intensity of fluorescence of annexin V.

Human Immunodeficiency Virus (HIV) infection is characterized by a depletion of the CD4+ T-lymphocytes. Absolute counts of CD3+CD4+ cells have been used in monitoring the progression of this disease and in assessing clinical trials of drugs developed to treat HIV infection and related complications [6,10]. In 1993, the definition of AIDS was revised to include HIV-infected persons whose CD4+ Tlymphocytes were ≤ 200 per μl, even in the absence of opportunistic infections or neoplasm. Both the CDC (Center for Disease Control) and WHO (World Health Organization) have emphasized the need to study lymphocyte subsets in normal populations [3]. It is thus necessary for a testing laboratory to establish normal ranges for these subsets in its local population. A number of studies have been done on lymphocyte subsets in healthy individuals. There is, however, a paucity of published data about Tunisians and North Africans in general. Therefore, our first objective in the present study was the establishment of normal ranges for CD3+CD4+ and CD3+CD8+ cells in a healthy Tunisian population. After that, the second step was the assessment of lymphocyte subsets in HIV infected individuals.

Resume Dans la presente etude, nous avons recherche les anticorps diriges contre le virus de l'hepatite C (VHC) chez 43 000 donneurs de sang volontaires de la population tunisienne par un test de depistage immuno-enzymatique. Nos resultats montrent que 0,7% (304/43 000) des donneurs ont un resultat de depistage anti-VHC positif. L'etude par immuno-blot des 304 serums depistes positifs a revele une prevalence confirmee de 0,18% (78/43 000) et a montre une variation du profil de la reponse immune en correlation avec les antigenes structuraux et non structuraux du virus de l'hepatite C: - Chez les donneurs dont les profils serologiques sont positifs (78/304) 25,6% : la reponse vis-a-vis de l'ensemble des antigenes (C+NS3 + NS4 + NS5) est la plus frequente 56,4% (44/78), la reponse vis-a-vis de deux antigenes se manifeste avec une frequence de 28,2% (22/78) et la reponse vis-a-vis de trois antigenes n'est que de 15,4% (12/78) avec une reactivite systematique vis-a-vis du « core . - Chez les donneurs dont les profils serologiques sont indetermines (99/304) 32,5% : la reponse vis-a-vis de l'antigene non structural NS5 est la plus frequente 54,5% (54/99), la reponse vis-a-vis de l'antigene non structural NS3 se manifeste avec une frequence de 27,3% (27/99) et la reponse vis-a-vis de l'antigene structural « core est de 18,2% (18/99), avec une absence d'anti-NS4.

The aim of this study was to access average delays for novogeneration of myeloid and lymphoid cells after allogeneic bone marrow transplantation (BMT) outcome and factors affecting this organization. A prospective analysis over 2 years (01/01/07 to 31/12/08) enrolling 19 children treated with allogeneic intrafamilial bone marrow transplantation. Indications for bone marrow transplantation were: aplastic anemia (3 cases), bemoglobinopathies (9 cases), myelodysplastic syndrome (1 case) and primary immunodeficiency (6 cases). Different conditioning regiments were used according to the indication. The study of immune reconstitution was based on the quantitative determination of immunoglobulin and lymphocyte subpopulation. These tests were routinely requested to 1 month, 2 months, 3 months, 6 months, 9 months and 12 months. The average time of engraftment was 18 days (12-24). A rate of CD4+T lymphocytes200/mm3 was provided within an average of 2,5 months (1-7). The average time to obtain CD8+T lymphocytes200/mm3 was 2 months (1-5). The humoral immune reconstitution was made within an average of 2 months (1-4). A report of CD4+/CD8+T lymphocytesI was obtained within 10 months and a half (1-24). Univaried analysis showed a correlation between the bone marrow sex matched and the faster reorganization of CD8+T cells (p=0.042). A quantity of CD34+6 10(6)/kg was significantly associated with the recapture of a formula lymphocyte CD4+/CD8+T1 (p=0.03) Immune recovery post bone marrow transplantation in children begins with myeloid lineage then lymphoid B then lymphoid T The inversion of the report CD4+/CD8+T lymphocytes, seems to be influenced by the high contain of CD34+cells in the graft as well as the type of conditioning.

Introduction Bone marrow (BM) represents the major source of mesenchymal stem cells (MSCs); however, umbilical cord blood (UCB) MSCs have some advantages over BM, such as a higher differentiation capability and noninvasive collection methods. Objectives We compared antigen expression and cytokine-secretion by MSC from BM and UCB expanded with media supplemented with fetal bovine serum (FBS) or human platelet lysate (HPL). Materials and Methods We compared protocols for the expansion of hMSC starting from samples of BM or UCB by morphological analysis, calculation of population doubling numbers, and cytometry techniques using monoclonal antibodies (BD Biosciences). Using the last technique, cytokines were detected in brain homogenate supernatant fluids of MSC cultured in various media, using the Bio-Plex cytokine assay system (BD Biosciences). Results Calculating the number population doubling (PD) and colony-forming unit-1fibroblast (CFU-F) assays showed significantly better expansion with HPL compared with a selected batch of FBS and within fewer days: PD about 5 for 10%HPL versus 25 for fibroblast growth factor2 (FGF2) medium. By flow cytometry, we observed a greater number of BM MSCs compared with UCB MSCs, as well as differences in the expression of some MSC antigens, particularly CD105, CD90, and CD31. Analysis of cytokines: FGFb, RANTES, VEGF, IL-6, IL-8, G-CSF, and GM-CSF showed only some of them to be expressed: namely, IL-6, IL-8, and VEGF. MSCs derived from UCB showed low concentrations of these cytokines compared with MSCs derived from BM.

Introduction Mesenchymal stem cells (MSCs) are multipotent cells able to differentiate into several mesenchymal lineages, classically derived from bone marrow (BM) but potentially from umbilical cord blood (UCB). Although they are becoming a good tool for regenerative medicine, they usually need to be expanded in fetal bovine serum (FBS)-supplemented media. Human platelet lysate (HPL) has recently been proposed as substitute for safety reasons, but it is not yet clear how this supplement influences the properties of expanded MSCs. Methods In the present study, we compared the effect of various media combining autologous HPL with or without FBS on phenotypic, proliferative and functional (differentiation, cytokine secretion profile) characteristics of human BM-derived MSCs. Results Despite less expression of adipogenic and osteogenic markers, MSCs cultured in HPL-supplemented media fully differentiated along osteoblastic, adipogenic, chondrogenic and vascular smooth muscle lineages. The analyses of particular specific proteins expressed during osteogenic differentiation (calcium-sensing receptor (CaSR) and parathormone receptor (PTHR)) showed their decrease at D0 before any induction for MSC cultured with HPL mostly at high percentage (10%HPL). The cytokine dosage showed a clear increase of proliferation capacity and interleukin (IL)-6 and IL-8 secretion. Conclusions This study shows that MSCs can be expanded in media supplemented with HPL that can totally replace FBS. HPL-supplemented media not only preserves their phenotype as well as their differentiation capacity, but also shortens culture time by increasing their growth rate.

The polymorphism of C4A and C4B genes was investigated in Tunisian patients with insulin dependent diabetes (IDDM) and compared to family members (sibs) and to healthy controls. Multiplex families were analysed. A significant increase in C4AQO (26.86% vs 6.90%) and C4BQO (40.29% vs 8.28%) phenotypes was noted in IDDM patients compared with controls. Using RFLP analysis, we confirmed the high frequency of C4 null alleles. We also observed that most of these alleles were genes deleted in IDDM patients (72.23% vs 20% for CA4QO and 74.07% vs 16.70% for C4BQO). A significant decrease in the C4B long (14.92% vs 67.12%) form of the gene was also demonstrated by RFLP analysis compared with controls. Two haplotypes were frequently associated with IDDM patients in whom the C4A and C4B were deleted genes.

The precision of immunological characterization of leukemias was improved by a certain number of technical innovations, particularly hybridoma production and standardization, resulting in monoclonal antibodies and definition of recognised cellular antigens (designated by CD: Cluster of Differentiation).The aim of this work was to determine the immunophenotyping profile of patients with leukemia, by means of a flow cytometric method: 66 blood samples coming from leukemic persons in the Sahel region were studied by flow cytometry, using about thirty monoclonal antibodies all marked with a fluorochrome, in one or two colour systems to assess their distribution according to type (lymphoid B or T / myeloid) and age, and to search for possible co-expressions of markers of different lineages.The marked preponderance of childhood B-ALL in our series is, at least partly, attributable to the age distribution of the Tunisian population. In agreement with studies from other countries, the majority of AML cases occurred among adults. A high proportion of AML cases in our series co-expressed markers of other lineages. Overall, accurate classification of acute leukemias was possible from a simple peripheral blood sample in 62 of 66 cases (93.9%).

To study by flow cytometry (FCM) the ploidy and the cellular cycle of nodular hidradenoma (NH) and hidradenocarcinoma (HC) and to assess the prognostic utility of this technique in such tumors.We studied retrospectively 2 HC and 11 NH one of which was considered as an atypical NH. Monoparametric study by FCM was realized on paraffin-embedded material. The extracted cells were marked by Propidium's lodure and cellular cycle was analyzed by the software Mod-Fit LT.Our study showed eleven 100% diploid profiles, 10 of which had low S-phase varying between 2 and 12%. All of these 11 tumors were NH. S-phase was high (23.79%) in a single case that corresponded to the atypical NH. Two tumors showed aneuploid profiles; these corresponded to the 2 HC.The results of the cytometric study suit perfectly to those of the histopathologic examination. FCM could so help to establish the prognosis of these tumors. But further studies are necessary to determine the value of this technique.

Apoptosis represents a particular form of programmed cell death which appears in all the damaged cells and potentially hazardous. It plays a crucial role in the development of multicellular organisms by assuring and maintaining the cellular homeostasis. Thus, apoptosis intervenes not only in the normal process of organisms' development but also in immune defence and in cancerous cells detection. Indeed, any blockage in the program of the apoptotic machinery would be responsible of some neurodegenerative and auto-immune diseases and could play a crucial role in different steps of carcinogenesis. Some researchers were very interested in studying apoptosis in hematopoietic stem cells CD34+ which could be intended to be reinfused to patients suffering from malignant diseases. They have noted that kinetic study of apoptosis of the hematopoietic stem cells CD34+ after the process of cryoconservation is also necessary. Such study permits to quantify the real and exact number of the viable hematopoietic stem cells CD34+ and therefore to eliminate such risk which would be associated with the reinfusion of apoptotic cells to patients. In this paper, we describe our contribution to hematopoietic stem cells CD34+ study by flow cytometry before and after cryopreservation by using annexin V as a specific probe allowing detection of phosphatidyl serine, one of the major features of apoptosis. But, we have noted a pronounced induction of apoptosis in peripheral mobilized blood compared to cytapheresis (after cryopreservation: 29.79% of apoptotic HSC CD34+ in peripheral mobilized blood but only 11.67% apoptotic HSC CD34+ in cytapheresis). Besides, we have noticed that hematopoietic stem cells CD34+ have had a statute of viability better than other mononuclear cells. These results put in value the reliability, the simplicity and the efficiency of flow cytometry for the analysis of apoptosis in hematopoietic stem cells CD34+ by following the intensity of fluorescence of annexin V.

Clinico-pathological study of superficial bladder cancer (pTa/pT1) informs about prognostic factors such as the size of the tumor, its uni or multifocal character, its grade and stage. Presently, these factors constitute the basis of therapeutic decision but do not allow to foresee the prognosis with certainty. Many technics have contributed to a better knowledge of such tumors; however, they have not allowed to fully master prognostic uncertainties. During the last decades, cell cycle and DNA content study by flow cytometry has been developping, bringing an additional prognostic element to various types of tumors. We have decided to study the impact of this technique to the assessment of the prognosis of superficial bladder tumors. The study concerned 65 patients presenting superficial bladder tumors (pTa/ pT1), with a follow-up of at least two years in case of not recurrence and in case of recurrence, having had a second resection with analysis of sections. Flow cytometry was applied to formol-fixed and paraffin-embedded endoscopic resection material of initial tumors by simple-labelling of DNA with propidium iodide. Following cytometric study, 35 (54%) of tumors were aneuploid and 30 (46 %) were diploid. For the diploid ones, S-phase mean value was 14.94% (from 2.72% to 33.43%); and G2M mean value was 8.3 (from 1% to 18%). The presence of an DNA- aneuploid peak had a predictive value of recurrence and progression in stage, with relative risks of 12 and 6.85 respectively. It was also correlated with the histological grade and stage. On the other hand, S-phase and G2M values had no prognostic significance.

A randomised crossover trial of two separators was undertaken to compare the mononuclear cell, CD34(+) cell and CFU-GM yield, in patients (61 years) with previously untreated symptomatic multiple myeloma. After first-line therapy, all patients received mobilising chemotherapy (cyclophosphamide 4 g/m(2)) and daily G-CSF. The first leucapheresis was performed on the first day the peripheral blood absolute CD34(+) cell count was20 cells/microl. All patients underwent 2 leucaphereses on consecutive days. The patients were randomised to undergo either the first or second leucapheresis using the COBE Spectra. The target duration of the procedure on the COBE Spectra was 2 total blood volumes, and for the Haemonetics MCS(+) it was 20 cycles with four recirculations. Between September 2003 and March 2005, 60 patients were entered in the study. COBE Spectra version 6 processed significantly larger volumes of blood than the Haemonetics MCS(+) (8,845 and 5,680 ml, respectively, P0.01). The absolute yield of mononuclear cells (2.1 vs. 1.5 x 10(8)/kg, P = 0.04), CFU-GM (11 vs. 3 x 10(4)/kg, P = 0.01) and CD34(+) cells (3 vs. 1.7 x 10(6)/kg, P = 0.02) were all significantly higher with the COBE Spectra version 6, as were the yields per unit volume of blood processed. In conclusion, our study shows that COBE Spectra Version 6 is faster and has a better yield than the Haemonetics MCS(+), in patients with multiple myeloma.

The biallelic NA antigen system is of special interest as the NA antigens are frequent targets of neutrophil antibodies, causing alloimmune neonatal neutropenia, blood transfusion reactions, and chronic benign autoimmune neutropenia in infancy. Neutrophils isolated from the peripheral blood of 119 unrelated individuals at the National Blood Center were phenotyped for NA1 and NA2 using a granulocyte immunofluorescence assay. A subsequent analysis of the phenotyping study showed that the NA1 and NA2 antigen frequencies were 0.529 and 0.865 respectively, and that the estimated NA1 and NA2 gene frequencies were 0.313 and 0.632 respectively. In conclusion, it was determined that the Tunisian population is of Caucasian origin. However, to validate this finding, further investigations are necessary.

A human recombinant erythropoietin is produced by transfection of a human EBV immortalised lymphoblastoid cell line by human erythropoietin gene. This cell line was selected because it is eucaryotic, human, able to grow in suspension and neither clonogenic nor tumorigenic. Purification of erythropoietin from culture supernatants of lymphoblastoid cells was described and structural, biochemical and functional characteristics were investigated in several studies. Results obtained for this kind of recombinant erythropoietin were similar to those obtained regarding the protein expressed in CHO cells suggesting that both molecules have the same efficiency in clinical applications.

Among the neutrophil polynuclear specific antigens (NA, NB, ND, NE, ...), NA antigen is the most common. It is a glycoprotein situated on the neutrophils FcRIIIb-receptor and presents 2 forms: NA1 and NA2. The epitope responsible of that polymorphism has got an amino acids composition that is unknown. The first techniques used for their analysis were the microagglutination and the granulocytotoxicity-later, the immunofluorescence, the chemiluminescence and the MAIGA (Monoclonal Antibody Immobilized granulocyte Antigen) were introduced. These last years, more efficient techniques appeared like Flow Cytometry and Polymerase Chain Reaction (PCR) that allowed phenotyping and genotyping of neutrophil polymorphonuclear specific antigens. The studies indicated that NA antigen frequency varies according to the populations and the ethnics. NA2 allelic form is more frequent than NA1 in the caucasian population (88% VS 46%). In human pathology, NA antigen is implicated in the physiopathological mechanisms of the alloimmune and probably auto-immune neutropenies.

Antibodies to Hepatitis C virus (HCV) were tested in 43000 Tunisian blood donors by using enzyme immuno-assay. Our results show that 0.7% (304/43000) were anti-HVC positive. Of these 304.78 were confirmed anti-HCV positive (0.18%) by immuno-blot, and 99 displayed an indeterminate profile. Different immune responses were observed: In donors with positive serologic pattern (78/304), 25.6% response towards whole antigens (C + NS3 + NS4 + NS5) was frequently observed (44/78) 56.4%. Reactivity to 2 antigens was observed in 28.2% (22/78) and with 3 antigens in only 15.4% (12/78), with systematic reactivity to core. In donors with indeterminate serologic pattern (99/304) 32.5%, reactivity to non-structural antigen NS5 was the most frequently observed (54/99) 54.5%, reactivity to non-structural NS3 antigen was noted in 27.3% (27/99) and to core antigen in 18.2% (18/99). No donors with isolated reactivity to NS4 were observed in our series.

Haplotypes including HLA A, B, C, DR, and DQ were compared in a study population comprising 18 Tunisian multiplex families with diabetic children. Eighty haplotypes found in IDDM patients were compared with 148 haplotypes present in healthy family members. RFLP analysis showed that two DR subtypes were significantly more common in the diabetic haplotypes (DR4-DQw8: 82 per cent in IDDM members compared to 0 per cent in healthy members, p less than 0.001 and DR-Dw25: 56 per cent in IDDM patients compared to 16.7 per cent in healthy members, p less than 0.001) and these were in most cases found in haplotype combinations with HLA A2 B44 DR4 DRw53 and HLA A 24 B18 DR3 genes, respectively.

The presence of hepatitis B virus DNA, delta antigen and anti-delta antibodies was examined in 159 Tunisian chronic HBs Ag carriers: 45 were asymptomatic and 114 suffered from cirrhosis. Serum hepatitis B virus DNA was detected in two (4.5%) asymptomatic HBs Ag carriers and in 11 (10%) HBs Ag positive cirrhosis patients. The prevalence of HDV infection determined by the presence of anti-delta was relatively high in asymptomatic HBs Ag carriers (33%) and in HBs Ag positive cirrhosis patients (21%). Active ongoing HDV infection, detected by serum HD Ag and anti-delta IgM, was shown in five patients with cirrhosis and active hepatitis B virus replication. We conclude that hepatitis delta virus may be endemic in Tunisia and does not always inhibit hepatitis B virus replication.

The early detection of microalbuminuria in insulin dependent diabetes is considered as a sign of initial stage of nephropathy possibly reversible if glycemic balance is well maintained. This detection requires very accurate methods as radio-immuno-assays. Yet, they are so slow that they represent an obstacle to systemic detection. We report in this work an apparaisement of an immuno-nephelemetric and electro-immuno-diffusion of Laurell methods. Results reveal that immuno-nephelemetry and electro-immuno-diffusion are a sensitive and accurate methods (threshold of sensitivity of Laurell method is : 2.5 mg/l and nephelemetry is 1.5 mg/l). Moreover, RIA, immunonephelemetry and Laurell methods are significantly well correlated (r = 0.903: Laurell/Radio-immuno-assay, r = 0.907: Nephelemetry/Laurell). In conclusion immuno-nephelemetry and electro-immuno-diffusion of Laurell are a choice methods to test great lines of samples. Radio-immuno-assay can be used as a reference method of detection of microalbuminuria.

Background: Thalidomide which represents an effective treatment strategy for relapsed/refractory multiple myeloma, actually represents a standard of care also for newly diagnosed multiple myeloma patients. Methods: In the present study, we adopted a gene expression profiling (GEP) strategy in an attempt to predict response (> 50% reduction in serum M protein) to primary therapy with thalidomide-dexamethasone for newly diagnosed multiple myeloma. Plasma cells (CD138+) were purified from bone marrow aspirates from 17 patients at diagnosis, before initiation of treatment with thalidomide-dexamethasone. GEP was performed using the Affymetrix U133 Plus_2 microarray platform. The Affymetrix output (CEL files) was imported into Genespring 7.3 (Agilent technologies) microarray analysis software, where data files were normalized across chips using GCRMA and to the 50th percentile, followed by per gene normalization to median. Criteria of response were those established by Bladè et al. Results: After sufficient follow-up, responders (n=9) and nonresponders (n=8) were identified, and gene expression differences in baselines samples were examined. Of the 11000 genes surveyed, Wilcoxon rank sum test identified 149 genes that distinguished response from non response. A multivariate step-wise discriminant analysis (MSDA) revealed that 14 of the 149 genes could be used in a response predictor model (see table). Of interest, the gene list encompasses WXSC1, known to be involved in the chromosomal translocation t(4;14) (p16.3;q32.3) in multiple myeloma. Conclusion: These results could be the first step to adopt microfluidic cards, in an attempt to select at diagnosis patients who will respond favourably to a particular treatment strategy. List of 14 genes able to predict response to primary therapy with thalidomide-dexamethasone for newly diagnosed multiple myeloma. Gene ID Gene Name Chromosomal location 212771_at C10orf38 10p13 229874_x_at LOC400741 1p36.13 219690_at U2AF1L4 19q13.12 202207_at ARL7 2q37.1 243819_at GNG2 14q21 203753_at TCF4 18q21.1 235400_at FCRLM1 1q23.3 211474_s_at SERPINB6 6p25 226785_at ATP11C Xq27.1 215440_s_at BEXL1 Xq22.1–q22.3 209054_s_at WXSC1 4p16.3 227168_at FLJ25967 22p12.1 213355_at ST3GAL6 3q12.1 223218_s_at NFKBIZ 3p12–q12

1,400 sera taken from patients suspected of having autoimmune diseases and sent to the laboratory for determination of antinuclear antibodies, are tested by comparative indirect immunofluorescence on 2 subtrata; rat liver section and HEP/2 cells. The 143 positive sera on rat liver sections are also positive on HEP-2. In the 1,010 sera which are negative on rat liver sections, 165 are positive on HEP-2, 113 give a nuclear fluorescence, 26 give a cytoplasmic fluorescence and 26 give a nuclear and cytoplasmic fluorescence. Three positive sera were also used in immunofluorescence on another cells: VERO and MRC 5, as well as dual immunodiffusion versus thymic and splenic cell extracts and 76 p. cent of these sera were found positive with these techniques. This confirms the advantage of the use of HEP/2 cells in demonstrating autoantibodies, especially when they are not detected on rat liver sections, like the anticentromer antibodies. This substratum offers the advantage of detecting not only antibodies directed against nuclear antigens, but also those directed against cytoplasmic antigens.

There is no doubt that the autoimmune process in human disease depends on genetic factors. Varying associations were noticed between HLA DR and autoimmune disorders. The frequency of HLA-A-B and DR antigens as well as the Bf and C4 allotypes have been investigated in insulinodependant diabetes mellitus (IDDM) and compared to that of healthy controls in Tunisian population. An increase of A30, DR3, DR4, BfF1, C4AQ0 and C4BQ0 and decrease of B40, DR2, DR5 and DR6 were found in diabetes when compared to the value observation controls. The strongest association was noticed with HLA, DR3 and DR4. The prospective role of DR2 and DR5 antigens were also confirmed. Examination of HLA, Bf and C4 alleles. Two supratypes associated with IDDM have been observed among the Tunisian patients.

The frequency of HLA A, B, and DR antigens as well as the Bf and C4 allotypes have been investigated in insulin-dependent diabetes mellitus (IDDM) and compared to that of healthy controls in the Tunisian population. An increase of A30, DR3, DR4, BfF1, C4Ao, and C4Bo and decrease of B40, DR2, DR5, and DR6 were found in diabetics when compared to the value observed in controls. The strongest association was noticed with HLA DR3 and DR4. Heterozygotes DR3 DR4 were very frequent in diabetics: 24.2 per cent versus 3.6 per cent in controls (relative risk 7.72). The protective role of DR2 and DR5 antigens were also confirmed. No supratypes of HLA, Bf, and C4 alleles associated with IDDM have been observed among these Tunisian patients.

Tataouine (South Tunisia) has been subject to several viral hepatitis epidemics during these last years. Prospective study has been conducted to define the prevalence of HVB infection. It has examined 511 cases, the age of whom was between 1 month and 70 years, reported in groups through the OMS recommendations, taking into account the number of inhabitants in each area. The global prevalence was 6.2% for HBs Ag and 18.5% for HBs antibody. These results were not different from the frequencies observed within the Tunisian population in general. The analysis of this study according to these groups, shows that in three areas (Ras El Oued, Bir 30, Dhibet) 60 to 80% of cases have at least one of HVB marquers, whereas the prevalence in other areas (Rogba) was very weak. The geographic repartition of HVB infection corresponds approximatively to areas that have been the most infected by the last hepatitis epidemics. A second study has completed and confirmed the results of the first one, taking into consideration a more important number of cases from areas of strong and weak endemicity. 596 sera have been examined in Rogba which counts 2000 inhabitants and is a weak endemicity area. 528 sera have been examined in the three areas of strong endemicity: 199 in Bir 30, 201 in Dhibet and 128 in Ras El Oued, which count around 2000 inhabitants, too. The percentage of cases presenting at least one of HVB serological marquers reaches 84 in Dhibet, 70 in Ras El Oued, 50 in Bir 30 and 24 in Rogba. This HBs Ag frequency is 3.8% in Rogba, 15% in Bir 30, 24% in Ras El Oued and 26.3% in Dhibet. It seems that the HVB is the virus of hepatitis epidemics observed in Tataouine at least in the three strong endemicity areas.