Novel inflammatory markers in patients with severe COVID-19 and a pulmonary thrombotic event.

Venous thromboembolism (VTE), clinically manifested as deep vein thrombosis (DVT) or acute pulmonary embolism (PE), is the third most common acute cardiovascular syndrome following myocardial infarction and stroke. The annual incidence of PE is between 39 and 115 per 100,000 inhabitants. The incidence of VTE is almost eight times higher in people aged 80 and older than in the fifth decade of life. We performed a retrospective study of 226 COVID-19 patients and selected group of patients who experienced a pulmonary thrombotic event. The incidence of PE in hospitalized COVID-19 patients was approximately 1.9-8.9%. The retrospective nature of the analyzed cohorts and relatively short observation periods could have led to underestimation of the actual incidence of PE. This study underlines the role of novel inflammatory biomarkers such as neutrophil to lymphocyte ratio and platelet to lymphocyte ratio in patients with a pulmonary thrombotic event in COVID-19. We suggest that these biomarkers may have high assessment value and complement routinely used biomarkers.

Prostate Infiltration by Treg and Th17 Cells as an Immune Response to Propionibacterium acnes Infection in the Course of Benign Prostatic Hyperplasia and Prostate Cancer.

Benign prostatic hyperplasia (BPH) and prostate cancer (PCa) belong to the most frequent diseases in ageing men. It has been proposed that prostate chronic inflammation is a risk factor for the development of both BPH and PCa. However, potential stimuli that cause or maintain inflammation in the prostate gland are still poorly characterized. Bacterial infections seems to be one of the potential sources of prostatitis. Recent studies show that Propionibacterium acnes (P. acnes) is the most prevalent microorganism in the prostate gland and may be a predisposing factor for inflammation of prostatic tissue. It indicates that P. acnes may contribute to cancer development by enhancing proinflammatory responses, as well as by modifying the prostate extracellular environment. In this review, we discuss the potential role of P. acnes in the development of BPH and PCa and highlight the importance of regulatory T CD4(+)FoxP3(+) (Treg) and Th17 cells in response to P. acnes infection in the context of both prostate diseases.

Infiltrating Treg and Th17 Cells of the Prostate Hypertrophy Gland Associated with Propionibacterium Acnes Infection.

PURPOSE: The aim of the study was to evaluate prostate cell infiltration by CD4(+)IL-17(+) and Treg cells in BPH and PCa patients depending on P. acnes infection in the prostate gland. PATIENTS AND METHODS: Prostate fragments were collected from 54 patients with PCa and 34 patients with BPH. Rapid ID 32 was used to identify the bacteria. Cells were analyzed by flow cytometry BD FACSCanto II. Statistical analysis was performed using Statistica 7 software (TIBCO Software Inc, USA). RESULTS: P. acnes was detected in 35% of patients with PCa and 41% of individuals with BPH. The infiltration of CD4(+)IL-17(+) and Treg cells was statistically significantly higher (P = 0.001) in patients with BPH and positive for P. acnes. A statistically considerably higher (P = 0.001) infiltration of Treg cells in treated for PCa with P.acnes infection was also demonstrated. CONCLUSION: Prostatitis caused by P. acnes may contribute to the development of BPH and PCa.

Changes of post-operative peripheral blood dendritic cells in patients undergoing laparoscopy.

INTRODUCTION: Surgical intervention affects local and systemic immune responses, especially in obese individuals. Many studies have attempted to evaluate immunological response to surgical trauma. Surgery changes the quantity and phenotype of circulating blood dendritic cells (DCs), including a decrease of total DCs post-operatively. The study aimed to evaluate the percentage and changes of myeloid, lymphoid DCs, and myeloid to lymphoid DCs ratio in obese and normal weight patients undergoing laparoscopy. MATERIAL AND METHODS: The study enrolled asymptomatic patients with gallstones, who underwent laparoscopic cholecystectomy. Blood samples were obtained before the surgery as well as 24 and 48 hours after the surgery. Cells were collected using a FACSCalibur flow cytometry, and phenotypes were analyzed with CellQuest software. RESULTS: No statistically significant differences were observed between obese and normal-weighted patients in all studied time periods, except for the myeloid to lymphoid DCs ratio assessed at 48-post-operative hour. The myeloid DCs percentage increased significantly in the post-operative period within both studied groups. The percentage of lymphoid DCs increased significantly in obese patients in all studied time periods. CONCLUSIONS: Laparoscopy induces immunomodulation, such as changes of myeloid and lymphoid dendritic cells, especially in obese patients. We describe new findings, in which minimally invasive surgical trauma promotes the increase of percentage of circulating DCs in the early post-operative period.

Immune system disturbances in Clouston syndrome.

BACKGROUND: Clouston syndrome belongs to the family of ectodermal dysplasias. So far, a defective immune response has not been reported in Clouston syndrome. We report, for the first time, immunological particularities of a large multigenerational Polish family with Clouston syndrome. METHODS: Five members of the same family with Clouston syndrome, aged 6-76 years, and 20 healthy volunteers, aged 19-73 years, were enrolled in the study. In all participants, the ability of neutrophils to phagocytize opsonized Escherichia coli was assessed. Granulocyte oxidative burst was determined quantitatively, and an isolation of peripheral blood mononuclear cells and the detection of lymphocyte subsets were performed. All patients with Clouston syndrome underwent microscopic assessment of hair shafts, x-rays of the skull and hand bones, extra- and intraoral examination, and panoramic x-rays. RESULTS: Compared to the controls, all patients with Clouston syndrome presented with significantly reduced phagocytic activities of granulocytes and monocytes (P < 0.05). The percentages of granulocytes and monocytes being positive for oxidative burst were also significantly reduced in all patients with Clouston syndrome (P < 0.05). No disturbances in the percentages and absolute counts of T CD3+, T CD3+/CD4+, T CD3+/CD8+, natural killer, and B CD19+ cells were found. CONCLUSION: Although this study expands knowledge about Clouston syndrome, it also raises many questions. The results provide evidence of significantly reduced phagocytic activity and oxidative bursts of cells playing crucial roles in a nonspecific immune response. Further studies are required to understand the underlying mechanism of the hereby described abnormalities.

Immunomodelling Characteristics of Mature Dendritic Cells Stimulated by Colon Cancer Cells Lysates.

UNLABELLED: Application of cells with high TAA (tumor associated antigen) presentation potential seems to be crucial in neoplasia immunotherapy. Such feature is distributed in dendritic cells, which present peptides from processed TAA - MHC molecules complex to the T cells of a host. The aim of the study was to assess the influence of colon neoplasia tissue lysate on functioning of generated autologous DC's in the field of autologous CD4+ lymphocytes immunological response towards Th1/Th2 under in vitro environment together with comparison and assessment of DCs' immunosuppressive properties acquired from patients with colon cancer. MATERIAL AND METHODS: The population of this study consisted of 16 healthy- controls, 36colon cancer patients. Blood samples were collected 24h before planned surgery and preventive antibiotic therapy. Neoplastic tissue sample, was digested for cell lysates preparation. DC's generation from PBMC was carried out in standard conditionsand medium enriched with rhGM-CSF and rhIL-4. Mature DC`s and cocultured autologous CD4 lymphocytes immunophenotype assessment was analyzed with flow cytometer. Intracellular and culture medium cytokines concentration was analyzed with ELISA and FACS method. RESULTS: DC`s generated from colon cancer patients stimulated with lysates presented greater maturity, lower expression of CD206 antigen, significantly higher expression of HLA-DR, CD208 and CD209 and high intracellular expression of IL-12, compared to non-stimulated cells. CONCLUSIONS: The neoplastic tissue in vivo produces a number of substances having an unfavorable effect on immune system, our results suggests using lysates as good dendritic cells stimulators that possibly could have application in colon cancer immunotherapy.

Evaluation of lymphocytes CD4+ and CD8+ and expression of ZAP-70 kinase on CD3+ and CD19+ lymphocytes in obese patients undergoing laparoscopic cholecystectomy.

BACKGROUND: Obesity has become a global epidemic and a leading metabolic disease in the world. Laparoscopic surgeries may influence the function of the immunologic system. The percentages of CD4+ and CD8+ T lymphocyte cells have been described as prognostic factors for patients undergoing abdominal surgeries. This study aimed to evaluate the changes in CD4+ and CD8+ T lymphocyte cells, the ratio of CD4+ to CD8+ cells, and the ZAP-70 kinase expression on T CD3+ and B CD19+ cells in obese and normal-weight individuals undergoing laparoscopic cholecystectomy (LC). METHODS: The study group consisted of 46 asymptomatic patients with gallstones shown by ultrasound examination but without signs of any gallbladder complications. The patients underwent planned LC. Blood samples were obtained at three times, and the percentages of studied cells were measured by flow cytometry. Patients were enrolled to two groups: N group (body mass index [BMI], ≤ 25 kg/m(2)) and O group (BMI, ≥ 30 kg/m(2)). For statistical analysis, the Mann-Whitney U test and the Wilcoxon matched-pairs signed-ranks test were used. All p values lower than 0.05 were considered significant. RESULTS: The percentage of CD4+ T cells did not differ between the N and O groups before or after the surgery. Only in the N group did the percentage of CD4+ lymphocytes increase from 0 to 48 h. A higher percentage of CD8+ lymphocytes was observed in the O group postoperatively than in the N group. Differences of ZAP-70 kinase expression in the O group were observed at 24 and 48 h of the study. Decreased expression of ZAP-70 kinase was shown in the N group at both 0-24 and 24-48 h. In the O group, this tendency was noted at 24-48 h. CONCLUSIONS: Immunologic activation after LC was confirmed in both weight groups. However, higher modulation, more typical for open surgeries, was observed in the obese group.

Evaluation of immature monocyte-derived dendritic cells generated from patients with colorectal cancer.

UNLABELLED: Dendritic cells are heterogeneous population of the leukocytes and most potent APC in activation of naive T lymphocytes. Therefore the DCs generated in vitro are under research for their application in anti-tumor immunotherapy. The aim of the study was generation of the immature dendritic cells from peripheral blood monocytes collected from colorectal cancer patients and comparison of their ability to endocytosis, cytokine production and immunophenotype to DCs generated from healthy donors. MATERIAL AND METHODS: 16 adenocarcinoma stage II patients were included in the study. Dendritic cells were generated in the presence of rhGM-CSF and IL-4. PBMC were isolated from the blood of patients and 16 healthy donors - control group. Immunophenotype, ability of endocytosis of Dextran- FITC as well as intracellular IL-12 expression of the generated dendritic cells was measured using flow cytometry. The cytokines (IL-6, IL-10, IL-12p70, IFN-γ) concentration in the supernatants of DCs culture was measured by ELISA. RESULTS: The percentage of the immature dendritic cells and expression of CD206 and CD209 antigens was significantly higher in patients group (p <0.05 and p <0.001 respectively). Significantly (p <0.001) higher expression of the antigens which initiate the Th2 immune response (CD80-/CD86 + and B7-H2 + / CD209 +) was in the patients group. There were no differences in endocytosis ability and the cytokines (IL-6, IL-10, IL-12p70, IFN-γ) concentration between investigated groups. CONCLUSIONS: High immature markers expression on the generated dendritic cells together with identical endocytosis ability in patients group is advantageous in antitumor autologous cells immunotherapy planning. However there is one troubling fact--high expression of markers, which may induce tolerance to particular antigen. It seems to be more reasonable to use the autologous DCs in the antitumor immunotherapy, especially due to the incompatibility in allogenic cells in the context of HLA complex.

Transdermal delivery of 8-methoxypsoralene mediated by polyamidoamine dendrimer G2.5 and G3.5--in vitro and in vivo study.

In this work, we have focused on 8-methoxypsoralene (8-MOP) complexed with G2.5 and G3.5 poly(amido amine) (PAMAM) dendrimers. The purpose of this study was to investigate the efficacy of half-generation G2.5 and G3.5 PAMAM dendrimers conjugated with 8-MOP for delivery of 8-MOP in vitro study through polivinyldifluoride membrane (PVDE) and prepared pig ear skin (PES) using Franz diffusion and in vivo study through the skin of experimental animals (hairless rat skin). The tissue concentration of 8-MOP in hairless rat skin was analyzed by high performance liquid chromatography (HPLC) after 1 and 2 h. Detailed distribution of 8-MOP in skin layers and cellular structures were analyzed using laser scanning microscopy (CLSM). In vitro and in vivo studies showed that half-generation G2.5 and G3.5 PAMAM dendrimers are able to facilitate transdermal delivery of 8-MOP. G2.5 PAMAM dendrimer appeared to be more effective 8-MOP penetration enhancer than G3.5 PAMAM dendrimer, but in vivo the differences are not statistically significant. The concept of using G2.5 and G3.5 PAMAM dendrimers as carriers seems to be a promising method for the delivery of 8-MOP for PUVA (psoralen-UV-A) therapy.

Biochemical markers of psoriasis as a metabolic disease.

Psoriasis is a chronic immune mediated inflammatory skin disease with a population prevalence of 2-3%. In recent years, psoriasis has been recognized as a systemic disease associated with metabolic syndrome or its components such as: obesity, insulin resistance, hypertension and atherogenic dyslipidemia. Many bioactive substances have appeared to be related to metabolic syndrome. Based on current literature, we here discuss the possible role of adiponectin, leptin, ghrelin, resistin, inflammatory cytokines, plasminogen activator inhibitor 1, uric acid, C-reactive protein and lipid abnormalities in psoriasis and in metabolic syndrome.

[The evaluation of apoptosis in peripheral blood lymphocytes in laryngeal cancer patients]. / Ocena apoptozy limfocytów krwi obwodowej u chorych leczonych z powodu raka krtani.

UNLABELLED: The laryngeal cancer is the most often cancer among others in head and neck region. It occurs mostly among 55 and 69. Its development depends on immunological state of the body. Vitality of the immunological system cells was considered due to growth, treatment sensitivity and prognosis of some neoplasms. The aim of this work were estimation and comparison the phenomenon of lymphocytes T and B apoptosis in laryngeal cancer patients treated with surgery and radiotherapy. MATERIAL AND METHODS: The material were 30 patients hospitalized in The Department of Otolaryngology Medical University of Lublin. They all were treated with surgery or surgery and radiotherapy. Apoptosis was estimated on different stages of the treatment process. All samples were examined with the flow cytometry method. The control group were 21 patients hospitalized because of the suspicion of the apnea syndrome, which wasn't confirmed with polysomnographic examination. RESULTS: Results of this study show significantly increasing percentage of peripheral blood apoptotic B (CD19+) cells caused by surgical treatment. The results considering radiotherapy showed different influence on the phenomenon of immunological cells apoptosis, still those results weren't significant. CONCLUSIONS: The surgical treatment causes increased amount of apoptotic peripheral blood lymphocytes.

Effect of polyamidoamine dendrimer G3 and G4 on skin permeation of 8-methoxypsoralene--in vivo study.

In the present study we have assessed the ability of (PAMAM) dendrimers G3 and G4 to facilitate transdermal delivery of 8-methoxypsoralen (8-MOP) in vivo. In vitro study using Franz diffusion cell revealed an enhanced transdermal flux for 8-MOP in complex with G3 and G4 dendrimer in relation to standard 8-MOP solution. In present study in vivo skin permeation potential of 8-MOP complex with G3 and G4 PAMAM dendrimer was assessed using confocal laser scanning microscopy (CLSM), which revealed an enhanced permeation of the 8-MOP to the deeper layers of the skin and significantly higher concentration in comparison with standard 8-MOP solution. Skin tissue 8-MOP concentration, evaluated by HPLC indicates that G3 and G4 PAMAM application significantly increase 8-MOP skin deposition in comparison with standard 8-MOP solutions after 1 and 2h. G4 appeared to be a more effective 8-MOP penetration enhancer than G3 PAMAM. Our results suggest the feasibility of G3 and G4 PAMAM dendrimers for transdermal delivery of 8-MOP resulting in better skin permeation and higher concentration of 8-MOP in epidermis and dermis of the drug that could help to improve effectiveness and safety of PUVA therapy.

Activated and inactivated PPARs-γ modulate experimentally induced colitis in rats.

BACKGROUND: This study sought to define the mechanism by which PPAR-γ ligands affect the course of experimentally induced colitis in rats. MATERIAL/METHODS: Inflammation was induced in Wistar rats by a single rectal administration of 2,4,6,-trinitrobenzene sulfonic acid (TNBS). The antagonist of PPARγ antagonist, bisphenol A diglycidyl ether (BADGE), was administrated intraperitoneally 120 mg/kg 4 times every other day. Rosiglitazone 8 mg/kg was administrated by gastric tube 4 times. Body weight was measured daily. After killing, the large intestinal tissue was weighed and collected for histopathologic and immunoenzymatic tests. Levels of IL-6, IL-10, and myeloperoxidase (MPO) were determined in serum and in intestinal homogenates. RESULTS: Rats receiving rosiglitazone had higher body weight, whereas large intestine weight/length ratio was lower; histology showed fewer inflammatory markers. Rats receiving TNBS and TNBS along with BADGE had more intensive inflammatory changes. Rosiglitazone alone decreased expression of IL-6; used with TNBS it decreased expression of MPO in intestinal tissue, yet did not increase the expression of IL-10. Decreased levels of MPO indicate reduced neutrophil-dependent immune response. The antagonist of PPAR-γ increased IL-6 in serum and decreased IL-10 in intestinal homogenates. Bisphenol A diglycidyl ether administrated to healthy animals increases serum IL-6 levels. CONCLUSIONS: Rosiglitazone inhibits experimental inflammation; administration of its selective antagonist abolishes this protective influence. Rosiglitazone inhibits expression of proinflammatory IL-6 and does not affect IL-10. Agonists of PPARs-γ are possibilities for inflammatory bowel disease prevention. Exogenous substances blocking PPARs-γ may contribute to development or relapse of nonspecific inflammatory bowel diseases.

Highly increased maspin expression corresponds with up-regulation of miR-21 in endometrial cancer: a preliminary report.

BACKGROUND: Maspin and programmed cell death 4 (Pdcd4) are tumor suppressor genes, and miR-21 is overexpressed in many solid tumors and was proven to negatively regulate a number of tumor suppressor genes including maspin and Pdcd4.The purpose of this study was to investigate the expression of maspin, Pdcd4, and miR-21 and their interrelations with clinicopathologic features in endometrial cancer using a quantitative approach. METHODS: Maspin, Pdcd4, and miR-21 expressions were evaluated by a real-time polymerase chain reaction in 20 endometrial cancer and 10 normal endometrium samples. RESULTS: Maspin showed a significantly increased expression in endometrial cancer samples compared with the control group and was up-regulated by a mean factor of 46.54 (SE range, 2.367-1160.26; 95% confidence interval, 0.515-15001, P < 0.0001). Expression of miR-21 was found significantly up-regulated in the sample group in comparison to control group by a mean factor of 2.312 (SE range, 0.741-7.778; 95% confidence interval 0.191-15.0, P = 0.028). No significant differences were present in the expression level of Pdcd4 between endometrial cancer and control groups. Comparison between IA and more advanced International Federation of Gynecology and Obstetrics stages of endometrial cancer in regard to expression levels of maspin, Pdcd4, and miR-21 did not reveal any significant differences. Similarly, no differences were encountered when histopathologic grading, myometrial invasion, age, body mass index, and parity were taken into consideration. CONCLUSIONS: Association between increased maspin expression and up-regulation of miR-21 in endometrial cancer suggests distinct and tissue-specific relationships of the 2 molecules in this type of malignancy and requires further studies that would reveal its clinical relevance.

Low atmospheric pressure and humidity are related with more frequent pulmonary embolism episodes in male patients.

Pulmonary embolism is a frequent condition, related with high mortality. Frequency of pulmonary embolism episodes has been related with several meteorological factors. The aim of the study was to analyze the influence of meteorological factors on the occurrence of pulmonary embolism in male and female patients. Medical data of patients hospitalized at our institution in 2007-2008 was analyzed. Study group included 400 patients with pulmonary embolism, living in the region located at an average level of about 200 m above sea level, with climate of a transitional character between maritime and continental climates. No significant differences in seasonal distribution of pulmonary embolism episodes were observed. A significant inverse correlation of the number of pulmonary embolism cases and atmospheric pressure, as well as air humidity, was identified in male patients. No significant correlations of temperature, wind velocity, precipitation and number of PE cases were observed. The influence of meteorological factors on the occurrence of pulmonary embolism in males is a new finding. A prospective study is warranted to further analyze this result.

Immunomodulatory effects of IFN-beta and lovastatin on immunophenotype of monocyte-derived dendritic cells in multiple sclerosis.

Multiple sclerosis (MS) is an autoimmune disease of the central nervous system and current MS treatment is only partially effective. Recent data suggest that statins may be potent immunomodulatory agents. In order to evaluate their role in MS, we analyzed the in vitro effects of interferon (IFN)-beta and lovastatin on the differentiation and maturation of monocyte-derived dendritic cells (DCs) of MS patients. Twenty-seven patients with relapsing-remitting MS were recruited for the study. DC differentiation and maturation were evaluated based on surface phenotypic changes and the expressions of CD14, CD83, CD1a, CD80, CD86, CD206, and C209 were analyzed by flow cytometry. The results showed that IFN-beta and lovastatin affect DC phenotype. Both agents decrease the expression of CD1a, which indicates a weakened presentation of glycolipid antigens. IFN-beta causes up-regulated and lovastatin down-regulated expression of CD86, which results in a biased Th-cell responses in MS. Furthermore, high doses of lovastatin cause a decrease in CD209 expression on the surface of DCs and can limit their migration to various tissues. One of the mechanisms of the beneficial action of IFN-beta and statins may be associated with their influence on DCs.

Selected metabolites of neutrophils in patients with 2-type diabetes complicated and non complicated with diabetic foot syndrome during colonization of E. coli toxin.

Neutrophils play an important role in the pathogenesis of complications of diabetes mellitus. The aim of the study was to evaluate the metabolism of neutrophiles activation markers during the colonization of E. coli endotoxin in order to determine their potential role in the treatment of 2-type diabetes complicated and non-complicated with the diabetic foot syndrome and to evaluate production of peroxide anions by stimulated and non-stimulated neutrophils depending on the exposition time. 54 patients were divided into 3 groups (15 healthy volunteers--control group (1), group 2 - 17 patients with 2-type non-complicated diabetes group 3 - 22 patients with diabetes and diabetic foot syndrome). Blood samples from all subjects were examined. Results show significant differences of E.coli endotoxin influence on metabolism of neutrophiles in study groups. Production of peroxide anions by non-stimulated neutrophils in 20th minute of the experiment was 15 times higher in the group with no diabetic foot and 18 times higher in the group with diabetic foot as compared to the control group. Production of peroxide anions produced by neutrophils increased significantly with the exposure time. The results correspond to data in the literature, that suggest, that type, time of exposition and concentration of pathogens may significantly interfere with neutrophiles activity in the course of diabetes.

SCID mice model in vivo evaluation of autologous and allogeneic dendritic cells activity on B-cell chronic lymphocytic leukemia.

In the present study we investigated in vivo therapeutic potential of DCs vaccines in B-cell chronic lymphocytic leukemia (B-CLL). On the day 0 the SCID mice were intraperitoneally inoculated with peripheral blood mononuclear cells (PBMC) of B-CLL patients at a dose of 10-30 x 10(6) and left untreated (controls) or i.p. injected on the day 7 with 0.2 - 14.0 x 10(6) dendritic cells. DCs were generated in vitro from peripheral blood monocytes of B-CLL donors (autologous DCs) or healthy donors (allogeneic cells) and pulsed with B-CLL antigens. On the day 14, the effect of implanted cells interactions was evaluated by a counting of CD19+CD5+ human leukemic cells and human T cells in the peritoneal fluid of mice. We found, that mean numbers of CD19+CD5+ leukemic cells as well as human T cells were lowered in peritoneal fluid of mice treated with allogeneic APCs. However, we did not observe similar effects with autologous DCs.

Immunotherapy with irradiated autologous leukemic cells in patients with B-CLL in early stages.

In this study 10(7) peripheral blood mononuclear cells including on average 78.56% CD19+/CD5+ lymphocytes were irradiated, and then administered intradermally as an anti-cancer vaccine to seventeen patients with B-cell chronic lymphocytic leukemia (B-CLL) at early stages (twelve injections, four at a weekly interval followed by eight vaccines given every two weeks). In eight out of seventeen patients, in the first two injections, irradiated leukemic cells were mixed with bacillus Calmette-Guerin (BCG) to improve the efficacy of therapy by additional induction of innate immunity. A hematological improvement (as defined by a >25% reduction of leukocyte count) to autologous leukemic cell vaccines was observed in 5/17 patients, stabilisation of disease in 5/17 patients and in 7/17 patients there was no response to immunotherapy. In seven patients significant increase of the lymphocyte doubling time was noted (p=0.02). There was no impact of BCG for immune responses or clinical outcome of vaccinated patients, but there was a significant increase of the absolute counts of CD3+ as well as of CD3+/CD4+ and CD3+/CD8+ T cells during the vaccination period. We observed a significant improvement of the phagocytic function of autologous dendritic cells generated from peripheral blood monocytes obtained from patients with B-CLL after the end of immunotherapy (p=0.006). An association between the clinical outcome and the percentage of leukemic cells positive for expression of ZAP-70 and CD38 was noted. In conclusion, our results demonstrated the feasibility and safety of autologous irradiated leukemic cell immunotherapy in patients with B-CLL. As we noted immunological, and to some extent, clinical responses, this approach merits further investigation, including the use of adjuvants other than BCG.