CORRELATION OF CD4+Т LYMPHOCYTES ACTIVATION WITH INTERLEUKIN IL-9, IL-17, IL- 22 PROFILES IN THE PERIPHERAL BLOOD OF PATIENTS WITH PLAQUE PSORIASIS.

Psoriasis is a T cell mediated chronic inflammatory skin disease affecting about 2% of the population worldwide. Recently has established the central role of IL-23/Th17 immune axis in the pathogenesis of psoriasis and different subclasses of T cells including Th1 and Th17 cells are involved in initiation and amplification of the skin inflammation process, in addition, in cases of recurrent psoriasis, Th22 cells play the role of memory cells with the help of Th9 cells, which are also important in this process. The main goal was to evaluate the ratio of T cell profile and IL23/Th17 axis by evaluating IL17A, IL22, IL9 in peripheral blood of persons with moderate to severe plaque psoriasis. We have estimated the activation of IL-23/Th17 axis by evaluating the level of IL-17A, IL-22 and IL-9 in peripheral blood of patients with plaque psoriasis (n=18) with different severity of the disease (PASI from 10 to 40) comparing the results with data obtained from healthy persons (n=15). The expression of CD69 activation marker on T helper cells has been evaluated as well. The results were analyzed using FACScan flow cytometer (Becton Dickinson). The percentage of CD3 + T lymphocytes in the peripheral blood of patients with psoriasis was not significantly different compared to normal healthy volunteers, however, the level of expression of CD4 + T cells was reduced. We observed a dramatic increase in IL22 along with a decrease in the level of expression of IL-9 and IL-17, the expression of Th activation marker (CD69) was also decreased in comparison with the control group. The T cell profile and the IL-23/Th17 axis functional activity levels were significantly different from the literature data obtained about the inflammatory region (psoriatic lesions on the skin). IL-9 and IL-17 expression levels are decreased in peripheral blood Th cells, which may be explained by mobilization of the corresponding Th9 and Th17 cells into the inflammatory site.

DIRECT-ACTING ANTIVIRALS FOR HEPATITIS C DO NOT AFFECT THE RISK OF DEVELOPMENT OR THE OUTCOME OF HEPATOCELLULAR CARCINOMA.

HCV infection and its complications, especially hepatocellular carcinoma, is a substantial public health burden. In 2015 "Nationwide hepatitis C elimination program" was launched in Georgia. According to the protocol, patients with HCC also receive DAA antiviral treatment. We study the effect of the different DAA therapy regiments on the incidence or recurrence of HCC and its prognosis. Overall, 408 patients were recruited in Georgian-French Joint Hepatology Clinic HEPA between April 2015-March 2016. The selection criteria were as follows: 1 - age 50-65 years; 2. Liver fibrosis level F3-F4 or cirrhosis at least 15 years of disease history; 3. HCV positive diagnosed by PCR method, whatever the level of viral load and genotype; 4. absence of previous complications of cirrhosis (ascites, gastrointestinal bleeding or HCC; 5. Child-Pugh class A or B; and 6. absence of severe extrahepatic disease. Essential clinical and biological parameters were recorded. Clinical monitoring and management of adverse events were performed on a regular base. HCV All patients included in the study received anti-HCV treatment with direct-acting antivirals (DAAs) within the national hepatitis C elimination program in accordance with national protocols. During April 2015-March 2016 treatment was provided with sofosbuvir (SOF) in combination with ribavirin (RBV), with or without pegylated interferon (IFN). Since March 2016, ledipasvir/sofosbuvir (LDV/SOF) was prescribed to all patients with or without RBV depending on the HCV genotype, level of fibrosis, and previous treatment experience. In conclusion, we find that neither different DAA regimens nor different treatment duration affects HCC risk after antiviral treatment. Moreover, there are no significant changes in mortality rate due to HCC in these groups. Therefore, it can be concluded, that HCC status is not a contraindication for DAA treatment, especially at the early stages of cancer, when a tumor is curative.

ATTACHMENT OF CHRONIC LYMPHOCYTIC LEUKAEMIA CELLS BY AUTOLOGOUS POLYMORPHONUCLEAR NEUTROPHILS MEDIATED BY BISPECIFIC ANTI-CD19/CD64 ANTIBODY.

Chronic lymphocytic leukemia (CLL) is the most common adult leukaemia in the US and in Europe, including Georgia. CLL presents with clonal expansion and accumulation of CD5+CD19+CD23+ cells in peripheral lymphoid organs and tissues and in bone marrow. The disease remains incurable, albeit there are new molecular and immunotherapy methods currently available, which, in conjunction with chemotherapy, lead to the "precision therapy" approach. The majority of immunotherapies are based on the ability of therapeutic antibodies to mobilize anti-tumour potential of immune responses. Bispecific antibodies (BsAb) are also considered in the treatment of CLL, whereby phagocytic cells play a key effector role in the destruction of the target CLL cells. Anti-CD19/anti-CD64 BsAb binds to CD19 receptors on CLL cells and to CD64 receptors (FcγRI) on monocytes and activated polymorphonuclear neutrophils (PMNs), thus inducing phagocytosis of the leukaemic cells. The aim of this study was to evaluate the ability of anti-CD19/CD64 BsAb to enhance adherence of CLL cells by PMNs, intact or activated with G-CSF and IFNγ cytokines. Membranes of the isolated CLL cells of 16 patients were stained with Red Fluoresent Linker and CLL cells were co-incubated with isolated autologous PMNs, intact or pre-stimulated with G-CSF and/or IFNγ for 4h or 24h. The PMN/CLL cell adhesion was analyzed with the FACScan flow cytometer by gating on PMNs with adhered RFL-stained CLL cells. The results were heterogenous. Our data demonstrate that anti-CD19/anti-CD64 BsAb has limited capacity to enhance leukemic cell attachment by autologous PMNs. This could partially be explained by the remarkable intensity of spontaneous ability of PMNs to adhere to autologous CLL cells. Pre-treatment of PMNs from CLL patients with G-CSF and INFγ, alone or jointly did not enhance the adhesion of the leukaemic cells. Moreover, G-CSF and IFNγ joint effect led to the reduction in the adhesion capacity of the effector PMNs. It appears, that therapeutic effect of anti-CD19/anti-CD64 BsAb on enhancing attachment of leukaemic cells to PMNs in CLL patients is limited and its application should be based on the assessment of individual capacity of the patients' phagocytic cells.

CHARACTERISTICS OF DIARRHEAL DISEASE COMPLICATED WITH HEMOLYTIC UREMIC SYNDROME AMONG CHILDREN IN GEORGIA, 2009-2016.

Shiga toxin-producing Escherichia coli (STEC) causes illness ranging from mild diarrhea to bloody diarrhea, to the hemolytic uremic syndrome (HUS), which manifests with a triad of microangiopathic hemolytic anemia, thrombocytopenia and acute renal failure. Surveillance of HUS and bloody diarrhea is not performed in Georgia. The primary objective of our study was to determine the annual incidence of diarrheal diseases. The secondary objective was to assess epidemiological characteristics, etiology and risk factors of diarrhea and HUS in Georgia among children under 15. We collected a retrospective data on diarrheal diseases particularly bloody diarrhea and HUS among children in Georgia in 2009-2016 years. Laboratory, clinical and epidemiological data was entered into electronic database. Descriptive statistics, proportions, incidence rates, means and medians were calculated in R statistical language using statistical package R for windows v 3.4.3. A total of 316 cases of bloody diarrhea including 64 (20.2%) cases complicated with HUS under age 15 were identified from 2009 until 2016. From 316 patients 5 (1.6%) have died, all of them with diagnosis and severe complications of HUS. Average rate of HUS per 100,000 populations during 2009-2016 was 0.3 and for bloody diarrhea 2 per 100,000. High RR for food products consumed by children with bloody diarrhea either complicated with HUS or not were various ice-creams produced locally (RR 4.23 P<0.001), dairy products (RR 2.79 P = 0.01), ground beef products (RR 4.52 P<0.001). The another highest attack ratio was identified for fruits (RR 6.19 P<0.001) and vegetables (RR 3.45 P < 0.001). Different enteric pathogens including shiga toxin producing E. coli was detected as etiology of diarrheal diseases and HUS. Epidemiological data suggests that inadequately washed fruits, vegetables and eating undercooked food and ice-cream could be a possible risk factors of exposure with enteric pathogens and developing diarrhea and HUS among children. Further investigations of food products are required to determine epidemiology and source food products of bloody diarrhea and HUS among children in Georgia.

Tularemia transmission to humans: a multifaceted surveillance approach.

Tularemia has sustained seroprevalence in Eurasia, with estimates as high as 15% in endemic regions. The purpose of this report is to characterise the current epidemiology of Francisella tularensis subspecies holarctica in Georgia. Three surveillance activities are summarised: (1) acute infections captured in Georgia's notifiable disease surveillance system, (2) infectious disease seroprevalence study of military volunteers, and (3) a study of seroprevalence and risk factors in endemic regions. Descriptive analyses of demographic, exposure and clinical factors were conducted for the surveillance studies; bivariate analyses were computed to identify risk factors of seropositivity using likelihood ratio χ2 tests or Fisher's exact tests. Of the 19 incident cases reported between 2014 and August 2017, 10 were confirmed and nine met the presumptive definition; the estimated annual incidence was 0.12/100 000. The first cases of tularemia in Western Georgia were reported. Seroprevalences of antibodies for F. tularensis were 2.0% for military volunteers and 5.0% for residents in endemic regions. Exposures correlated with seropositivity included work with hay and contact with multiple types of animals. Seroprevalence studies conducted periodically may enhance our understanding of tularemia in countries with dramatically underestimated incidence rates.

PHAGOCYTOSIS AND EXPRESSION OF FCg-RECEPTORS AND CD180 ON MONOCYTES IN CHRONIC LYMPHOCYTIC LEUKEMIA.

Chronic lymphocytic leukaemia (CLL) is a lymphoproliferative disease characterised by accumulation of monoclonal CD19+CD5+CD23+ lymphocytes in the peripheral blood and bone marrow. CLL is the most common type of the adult leukemia in the Western world. The disease is incurable, albeit there are new molecular and immunotherapy methods currently available in conjunction with chemotherapy, leading to the "precision therapy". The majority of immunotherapeutic approaches are based on the ability of therapeutic antibodies (Rituximab, Alemtuzumab) to mobilize anti-tumour potential of the Natural Killer cells and macrophages/monocytes through their Fcg-receptors (FcγR). Therefore functional status of monocytes in CLL is an important contributor to the efficacy of this treatment. In addition, CLL patients are characterized by a profound immunodeficiency, and how this affects monocytes, has not been established. Here we study ex vivo phagocytic function and the expression of Fcg receptors and CD180 toll-like receptor (TLR) by monocytes of 14 untreated and 8 treated with Cyclophosmamide, Adriamycin, Prednisolone (COP) CLL patients, and 12 age-matched control volunteers. Phagocytic function was assessed through the ability of freshly isolated monocytes to attach and to engulf intact or opsonized Staphylococcus aureus particles in vitro with or without Granulocyte/Macrophage Colony Stimulating Factor (GM-CSF) and Interferon g (IFNg). Simultaneously, immunophenotyping for FcγRI (CD64), FcγRII (CD32), FcγRIII (CD16) and CD180 has been carried out. The results were assessed by Flow Cytometry. Our results demonstrated that phagocytosis of the intact and opsonised or intact S. aureus by monocytes of CLL patients was significantly decreased in comparison with normal controls, with no recovery upon the treatment with GM-CSF and IFNg. A significant decrease in the expression of CD64 and CD180 has been detected on monocytes of CLL patients, with the drop in CD64 expression correlating with the disease progression and advanced Rai stages. In addition, the treatment with COP led to a more profound decrease in the expression of CD64. No appreciable changes were detected in the expression of CD32 and CD16 throughout the experiments. The diminished expression of CD64 and CD180 provides possible explanation for the impaired phagocytic function of monocytes in CLL, as FcγRI receptor interaction with opsonising IgG1, and CD180 with the ligands on S. aureus might affect the ability of monocytes to attach and to engulf S. aureus particles and effectively eliminate the pathogen. Our data indicates that the decreased functional status of monocytes in CLL might contribute to the diminished efficacy of therapeutic antibodies as well as to the immunodeficiency, characteristic for these patients.

PROSPECTIVE OBSERVATIONAL COHORT STUDY OF PREDICTORS OF THE EFFECTIVENESS OF PEGYLATED INTERFERON IN PATIENTS WITH HEPATITIS C IN GEORGIA.

The main objective of the study was to evaluate the predictive values of virological response at 4th and 12th weeks after treatment initiation on sustained virological response by HCV genotype in patients with hepatitis C in Georgia. Local, non-interventional, prospective, cohort study was conducted in 2011-2016. A cohort of adult peginterferon treatment naïve patients with chronic hepatitis C were observed during the complete active treatment period with PEGASYS®/COPEGUS® and 24 weeks after the end of treatment. HCV RNA titers were assessed prior to treatment, after 4, 12 weeks of treatment, at the end of treatment and 24 weeks after the end of treatment.530 men and women aged ≥18 years with serologically proven CHC (all genotypes) were enrolled in this study. Enrolles study subjects were treated with PEGASYS® 180mkg (peginterferon alfa-2a) in combination with COPEGUS® 200mg (ribavirin) according to the current standard of care and in line with current summary of product characteristics. All clinical information for this cohort study was collected from the patient's medical records. All laboratory parameters which were collected for this observational study according to the protocol were performed in study centers. All data were analyzed with descriptive and analytical statistics. Our analysis demonstrated that the early achievement of viral response predicts the higher probability of achieving sustainvirul response. The viral response itself was strongly associated with baseline liver fibrosis quantitive HCV RNA level. Early starting of treatment determines the probability of achievement higher osustain viral response.

MODULATION OF ANTITUMOR IMMUNE RESPONSE IN MOUSE PROSTATE CANCER MODEL.

Aim of study - prostate cancer is second most common cancer in men worldwide, and fifth leading cause of death from cancer in men (6.6% of the total men deaths). Unfortunately, once cancer spreads outside of prostate, it becomes incurable. Androgen deprivation can provide some relief, but resistance will develop eventually, and at that time no effective treatment options are left to the patient. Therefore, the search of alternative treatment modalities is paramount. In this article we evaluated the role of epigenetic modifier 5-azacitidine (5-AzaC) and immunomodulator - Lenalidomide and their possible impact on the immune response in the murine prostate cancer model. We studied their impact on murine prostate cancer cells and on dendritic cells (DC), the most potent antigen-presenting cells known. RM-1 is a murine prostate cancer cell line, resembling hormone-independent model, which is a lethal variant of this disease in humans. Dendritic cells were obtained from murine bone marrow. Cell proliferation assays were performed to evaluate the effect of 5-AzaC and lenalidomide on prostate cancer and DC. Flow cytometry, ELISA and real-time PCR was performed to evaluate the effect of these compounds on DC and prostate cancer cells. 5-AzaC treatment of RM-1 prostate cancer cells and DC resulted in the decreased proliferation for both, while lenalidomide had no effect. DC were treated with lenalidomide and the expression of surface markers MHC Class I, MHC Class II, CD80, CD86, CD 205, and CD40 was increased. Secretion of IL-12 and IL-15 by DC increased significantly with addition of 5-AzaC. There was also the change in the expression of endothelin receptors on DC, which can affect their function. 5-AzaC also resulted in the increased expression of cancer-testis antigen, P1A, by prostate cancer cells. Combination of epigenetic modifications and immunomodulation by 5-AzaC and lenalidomide should increase tumor immunogenicity and it enhanced DC function. Therefore, these compounds might have a role in the treatment of advanced prostate cancer.

COMPARATIVE STUDY OF FIB-4 INDEX AND TRANSIENT ELASTOGRAPHY AMONG PATIENTS WITH CHRONIC HEPATITIS C VIRUS INFECTION IN GEORGIA.

Liver biopsy remains the reference standard for fibrosis staging. However, it has several limitations, which have led to the development of non-invasive methods. We evaluated liver fibrosis severity among HCV infected patients by comparing transient elastography (TE) and FIB-4 index. Retrospective study was conducted. Clinical data for 750 patients were obtained. The mean age of the study population was 51 years; 595 (79.3%) were male and 155 (20.7%) were female. TE and tests on biological samples were performed within one-week timeframe. Additional analyses of prothrombin index, albumin concentration, splenomegaly on abdominal ultrasound and esophageal varices on upper gastrointestinal endoscopy were performed among selected patients. Comparable results were observed among 534 patients (71.2%). FIB-4<1.45 had a negative predictive value of 89% to exclude significant fibrosis and FIB-4>3.25 had a positive predictive value of 100 % to confirm the existence of significant fibrosis. Inconclusive FIB-4 score was obtained in 170 (22.7%) patients. Of them 127 (74.7%) had significant fibrosis (F3-F4) by TE. Discordant results (FIB-4 <1.45 and Liver Stiffness Measurement (LSM) >9.5 kpa) were observed in 46 (6.1%) of patients. Low prothrombin index, low albumin concentration, splenomegaly and esophageal varices were significantly (p<0.001) correlated with TE results. Discrepancy showing high FIB-4 score and low LSM was not observed in our cohort. There was a good correlation between TE and FIB-4 score. FIB-4 could rapidly replace expensive methods to assess liver fibrosis severity in some scenarios. However, our study demonstrated superiority of TE. LSM correlated better with indirect markers of significant fibrosis.

ASSESSMENT OF THE LEVEL OF KNOWLEDGE OF MEDICAL PERSONNEL IN DIARRHEA AND HEMOLYTIC-UREMIC SYNDROME.

Survey have been conducted among medical professionals to test knowledge level of HUS and diarrheal diseases and to identify predictor variables for better knowledge. Cross-sectional survey have been conducted among medical personnel at different clinics in Tbilisi and in regions of Georgia. Participants were selected from different clinics in Tbilisi and in three biggest regional cities (Zugdidi, Batumi and Kutaisi) of Georgia. A total of 12 clinics were selected from them 6 were in Tbilisi and 2 at each regional cities. Clinics were selected based on their ability to provide services for gastrointestinal diseases, infectious diseases and kidney diseases. Data were entered into electronic database and analyzed using R v3.3.2. Descriptive statistics and methods of multivariate analysis were used for data analysis. 366 medical personnel have been interviewed. 73% (267) were females and 27% (99) males. Mean age was 40.8, IQR (27-52). A total of 64% (235) participants were from clinics located in Tbilisi. In multivariate analysis background in infectious diseases, female sex and having more than 10 years of medical experience were significantly associated with the total knowledge score of diarrheal diseases (p<0.05). High total knowledge score of HUS was detected among pediatricians (p<0.05). Trainings has been recommended for medical specialists to increase knowledge of diarrheal diseases and HUS to be able to identify those condition and to provide timely medical support for patients.

INCIDENCE OF TUBERCULOSIS AMONG HIV/HCV CO-INFECTED PATIENTS RECEIVING HEPATITIS C TREATMENT WITH PEGYLATED INTERFERON AND RIBAVIRIN IN GEORGIA.

Treatment of hepatitis C is necessary for ensuring higher life expectancy among HIV/HCV co-infected patients. However antiviral treatment for chronic HCV infection with Pegylated interferon (PEG-IFN) and Ribavirin (RBV) is associated with a variety of side effects. In Georgia up to 22% of HIV-infected patients were found to have active Tuberculosis (TB) and 22.4 to 32.6% had latent TB. The objective of this study was to describe the characteristics and clinical outcomes of tuberculosis in HIV/HCV co-infected patients receiving hepatitis C treatment with pegylated interferon and ribavirin and calculate incidence rate of TB. A retrospective study was conducted among HIV/HCV co-infected patients receiving antiviral treatment for chronic HCV infection at the Infectious Diseases, AIDS and Clinical Immunology Research Center, Tbilisi, Georgia from December 2011 to May, 2015. A total of 420 HIV/HCV co-infected patients received HCV therapy with PEG-IFN and RBV during study period. Six of 420 patients developed TB while receiving PEG IFN + RBV therapy. These patients were on Antiretroviral treatment. Baseline HIV RNA load was <34 copies/ml and CD4+ cell counts >350 cells/mm3. No opportunistic infections were observed in all cases. Three of 6 patients had a previous positive tuberculin skin test (TST) result and had completed isoniazid chemoprophylaxis several years before TB diagnosis. In 2 patients TST was not performed. Only one patient had experienced a previous episode of TB and had completed the anti-TB therapy 1 year before hepatitis C treatment. In all patients TB was diagnosed during the PEG IFN + RBV therapy. Hepatitis C treatment was immediately stopped in all patients. The incidence rate of TB was 1.4 cases per 100 person-years (95% CI=0.58-2.97). Our study emphasizes the necessity of screening for latent TB prior to the initiation of chronic hepatitis C treatment with PEG IFN and RBV.

MOLECULAR TRACING OF HETEROSEXUAL HIV-1 TRANSMISSION IN GEORGIA.

HIV epidemic in Georgia has entered a new phase with number of heterosexually acquired infections rising each year. Epidemiological data indicates that this switch in epidemic trends is largely due to HIV positive male IDUs transmitting the virus to their female sexual partners. However, no genetic studies confirming linkage between IDUs and their sex partners were done in Georgia before. The objective of our study was to investigate molecular epidemiology of HIV-1 transmission events between heterosexual couples. Viral genotypes were obtained from plasma specimens of 36 heterosexual HIV-1 positive antiretroviral treatment (ART) naive persons representing 18 epidemiologically linked transmission events were genotyped and phylogenetic analyses were done on HIV pol sequences. HIV infection among all women was attributed to heterosexual transmission from their partners. None of 18 women had history of IDU. Fourteen pairs had subtype A virus, three - subtype B and one - subtype G viruses. Phylogenetic analysis confirmed the existing epidemiological link in 16 pairs with bootstrap values ranging from 88% to 100%. Of these 16 events, viruses from 14 pairs had genetic distance less than 0.015.Mutation A62V was seen in samples from 5 pairs, of them samples from 4 pairs additionally had V77I mutation. All 5 pairs were infected with the subtype Avirus. Women, who are sexual partners of IDUs or other men with high risk heterosexual behaviors, are at increased risk of HIV acquisition. HIV epidemic in Georgia has not spread to general population and remains concentrated around key populations at risk. Our work confirms that female sexual partners can serve as a bridge between key affected populations and general community, such as heterosexually active adults. Therefore, prevention efforts targeting key populations at risk and their sexual partners need to be expanded to avoid the spread of the infection within specific communities and beyond.

Distinct drug resistance profile of HIV-1 subtype A strain circulating in Georgia.

Emergence of HIV-1 drug resistance limits effectiveness of antiretroviral therapy (ART). Since 2004 Georgia provides free ART to all patients in need. We aimed to evaluate drug resistance patterns of Georgian HIV-1 variants among patients with virologic failure. Study included adult HIV-1 patients, who experienced virologic failure and were found to carry drug resistant strains based on genotypic resistance testing in 2005-2013. HIV-1 pol gene sequences were examined for the presence of resistance-associated mutations. Stanford HIV Sequence Database was used for interpretation of resistance data. A total 193 patients were included in the study. Among them majority (86.5%) carried subtype A virus and nearly 80% were on Efavirenz-based regimen. The most common nucleoside reverse transcriptase inhibitor (NRTI) mutation was M184V - 86.0% (n=166). The most frequent non-nucleoside reverse transcriptase inhibitor (NNRTI) mutation was G190S, found in 105 (54.4%) of samples. Other significant NNRTI mutations included K101E (31.6%, n=61), K103N (30.1%, n=58) and Y181CI (26.9%, n=52). The prevalence of G190S was 62.3% in subtype A viruses compared to 3.8% in non-A variants (p<0.0001). Frequency of K101E was also significantly higher in subtype A (36.5% vs. 0%, p<0.0001). ). In 69 samples G190S co-occurred with either K101E or Y181C or with both: 39 genotypes G190S/K101E; 10 genotypes G190S/Y181CI and 20 genotypes G190S/K101E/Y181CI. High prevalence of G190S and K101 mutations suggests subtype A specific response to currently approved first-line NNRTIs. Frequent co-occurrence of G190S with Y181C and K101E may limit the use of novel generation NNRTIs in subtype A infected patients with previous exposure to this drug class.

Correlation of the expression of CD32 and CD180 receptors on CLL cells and MEC1 cell line.

Chronic Lymphocytic Leukemia (CLL) presents with clonal expansion and accumulation of CD5+CD19+CD23+ cells in peripheral lymphoid organs and tissues and in bone marrow. CLL is supposedly driven by exogenous and/or endogenous (auto)antigen(s) and there is increasing evidence that CLL cells receive microenvironmental signals which support their growth, survival and expansion in vivo. We have previously shown that powerful signals are received by CLL cells through CD180 orphan toll-like receptor. Additional accessory signals could be generated through FcγRII (CD32), since both are expressed on CLL cells as well as on control B cells. Here we studied correlation of the expression of CD32 and CD180 on CLL cells as well as on MEC1 cell line. Peripheral blood mononuclear cells (PBMC) from CLL patients and age-matched healthy volunteers were separated, stained with appropriate antibodies to CD19, CD32 and CD180 and analysed by flow cytometry. CD32 and CD180 expression on MEC1 cells was studied at different time-points. The data was statistically analysed using the Mann-Whitney non-parametrical test. Our data indicates that expression of CD32 is significantly increased on CLL cells compared to control B cells as well as in long-term MEC1 cell culture. In contrast, CD180 expression on MEC1 cells significantly decreased throughout 0-96h of MEC1 cell culture. We have recently shown that CD180 ligation can redirect sIgM-mediated signaling from pro-survival to pro-apoptotic. This data indicates that a drop in the expression of CD180 on cycling CLL cells might lead to a weakening of this effect and enhance further survival and expansion of CLL cells in proliferative centres of lymphoid tissues. Since MEC1 cells are derived from a CLL patient with mutated IGVH genes (M-CLL) negative correlation between CD180 and CD32 expression on cycling MEC1 cells could be limited to M-CLL.

Aberrant expression of Fcγ-receptors and Toll like receptor CD180 on monocytes from patients with chronic lymphocytic leukaemia.

Chronic lymphocytic leukemia (CLL) is the most common leukaemia in the US and in Europe, including Georgia. Patients with CLL are susceptible to infectious diseases as a result of both, the disease progression and chemotherapy that indicates deficiency of immune responses to pathogens, including innate immunity, mediated by monocytes. Monocytes are also often recruited by monoclonal antibodies (mAbs) which express anti-tumour toxicity through Fcγ-receptor (FcγR)-mediated phagocytosis of opsonised leukaemic cells. In this paper we address of monocytes functional status through assessment of the patterns of expression of Fcγ receptors CD64, CD32, CD16 and CD180 receptor on monocytes from CLL patients and healthy individuals using specific mAbs and flow cytometry. Our data demonstrate that monocytes from peripheral blood of CLL patients lack expression of CD64 and CD16 as well as CD180 that would substantially undermine their ability to contribute to anti-bacterial immune responses. In addition, aberrant expression of CD64 would negatively affect the efficiency of antibody-mediated immunotherapies.

The cascade of care in the Eastern European country of Georgia.

OBJECTIVES: Individual and public health benefits of antiretroviral therapy (ART) rely on successful engagement of HIV-infected patients in care. We aimed to evaluate the HIV care continuum in the Eastern European country of Georgia. METHODS: The analysis included all adult (age ≥ 18 years) HIV-infected patients diagnosed in Georgia from January 1989 until June 2012. Data were extracted from the national HIV/AIDS database as of 1 October 2012. The following stages of the HIV care continuum were quantified: HIV infected, HIV diagnosed, linked to care, retained in care, eligible for ART and virologically suppressed. RESULTS: Of 3295 cumulative cases of adult HIV infection reported in Georgia, 2545 HIV-infected patients were known to be alive as of 1 October 2012, which is 52% of the estimated 4900 persons living with HIV in the country. Of the 2545 persons diagnosed with HIV infection, 2135 (84%) were linked to care and 1847 (73%) were retained in care. Of 1446 patients eligible for ART, 1273 (88%) were on treatment and 985 (77%) of them had a viral load <400 HIV-1 RNA copies/mL. Overall, 39% of those diagnosed and 20% of those infected had a suppressed viral load. CONCLUSIONS: The findings of our analysis demonstrate that the majority of patients diagnosed with HIV infection are retained in care. Loss of patients occurs at each step of the HIV care continuum, but the major gap is at the stage of HIV diagnosis. Reducing the number of persons living with undiagnosed HIV infection and simultaneously enhancing engagement in continuous care will be critical to achieve maximum individual and public health benefits of ART.

Discrepancy between HCV structural and non structural genes in Georgian genotype two patients.

Correct identification of hepatitis C genotypes is an important diagnostic tool, which guarantees further selection of adequate treatment regimen and correct duration. Ideal approach for accurate genotyping is amplification of both structural and non structural parts of HCV genome. As different methods, which use either one or another region for HCV genotyping sometimes lead to indeterminate genotype and subtype results. Therefore, it is of importance to compare HCV genotyping results based on two different genomic regions. As part of this study, remnant 108 specimens, with previous history of successful genotype identification by 5'UTR/core Versant HCV genotyping kit, were retrospectively analyzed. "In house" HCV real time PCR based method that amplifies parts of NS5B region was used for this purpose. Based on our data, genotyping calls were concordant between genotype one and genotype three specimens group in both regions. However, discrepancy was evident among genotype 2 group. Of 25 specimens originally typed as genotype 2 in structural region, only 7 was confirmed in non structural region, remaining 18 specimens were typed as 1b. Therefore, the discordance rate between structural and non structural regions for genotyping call among genotype two was 72%. Our data showed highly discordant structural and non structural genome for genotype two identification in our specimens. We propose that this phenomenon might be due to the recombination event between genotype two and genotype one. Possible circulation of this strain in Georgia stresses the need for detailed sequencing and phylogenetic analyses of these specimens in both structural and non structural parts of HCV genome.

Occurrence of occult HCV infection among Hiv infected patients in Georgia.

Occult hepatitis C (OCI) infection has been known as detectable HCV-RNA in the liver or peripheral blood mononuclear cells (PBMCs) in the absence of detectable serum or plasma HCV-RNA. OCI has been detected among different patients groups worldwide, it has been found not only in chronic hepatitis patients of unknown origin, but also among several groups at risk for HCV infection (hemodialysis patients or family members of patients with occult HCV). This occult infection has been reported also in healthy populations without evidence of liver disease. Prevalence of occult Hepatitis C virus has not been investigated in Georgian population, where a rate of HCV infection is highest (6.7%) among Eastern European Countries. The aim of this study was to investigate the occurrence of occult HCV infection among HIV infected individuals in Georgia. As a pilot study, we have selected three groups of HIV infected patients for analyses: Group 1- HIV infected patients without evidence of liver disease (n=98), group 2- HIV infected patients with cryptogenic liver disease (n=34) and group 3- HIV/HBV co infected patients (n=29). HCV RNA was tested in PBMCs samples by real-time polymerase chain reaction. HCV genotyping was performed by Line-probe assay based on reverse-hybridization technology. Liver fibrosis was evaluated by transient elastography (FibroScan®). HCV-RNA was detected in PBMCs specimens among 2 (2%) subjects from group 1, 4 (12%) subjects from group 2, and 9 (31%) subjects from group 3. HCV genotypes were determined for 14 of 15 OCI subjects resulting following genotype distribution: 6 (46%) - 1b, 3 (23%) - 2a/2c and 5 (38%) - 3a. One samples failed to be genotyped due to extremely low HCV viral load. Our data revealed the occurrence of occult HCV infection in HIV infected patients. No single HCV genotype was predominant in the present study. Liver fibrosis was found more frequently and the fibrosis score was significantly higher in OCI patients versus negative ones, suggesting that undiagnosed OCI might impact on the liver damage. The study demonstrated that testing only for HCV antibody fails to identify the true prevalence of HCV co-infection among HIV infected patients. We propose that in the absence of liver biopsy specimens, analysis of PBMC sample for HCV-RNA would be informative for detection of occult HCV.

[Results of monitoring of pollution by metallic impurities of environment of districts of anthropogenic impact in Georgia and methods to reduce their technogenic load].

There was performed a comprehensive study of consistent pattern of the accumulation of toxic heavy metals in the natural environments of mentioned areas and there were developed methods and recommendations for improvement of their ecological status. The aim of the study--a performance of the study of pollution of natural environments by major mining regions of Georgia: Madneuli copper pyrite and barite-polymetallic deposit, Chiatura manganese deposit, Lukhumi arsenic deposits. In the process of the work, the special attention was paid to the consistent patterns of accumulation of heavy toxic contaminants in natural waters and soils, as well as to the development of methods and recommendations for their purification from impurities. At the same time there were used: field geochemical, and hydrogeochemical pedological methods, modern ultrasensitive analytical equipment, laboratory experiments for the identification of tools of water and soil purification from heavy metals etc.

The first kidney transplant in HIV infected patient in Georgia.

Highly active antiretroviral therapy (HAART) has significantly improved the life expectancy of patients with HIV. As a result, kidney transplantation is considered an viable treatment option for HIV infected patients with end stage renal disease. The first living-related kidney transplant in Georgia has been performed between non-identical twin sisters in July 2013. In this paper we give the detailed case report and short overview of the existing literature. This is the first report of the successful kidney transplant in HIV infected patient in East Europe and former Soviet Union Countries including the Central Asia.