PSA-PSMA profiles and their impact on sera PSA levels and angiogenic activity in hyperplasia and human prostate cancer.

AIM: The relevance of prostate specific antigen (PSA)-prostate specific membrane antigen (PSMA) profiles in pathologic prostate (hyperplasia and cancer) has not been fully understood. The aim of this study is to investigate the impact of PSA-PSMA profiles on sera PSA levels and angiogenic activity in benign prostate hyperplasia (BPH) and prostate carcinoma (PC). PATIENTS AND METHODS: The study has been carried out in 6 normal prostate (NP), 29 BPH and 33 PC with dominant Gleason grade>8. Immunohistochemical analysis has been performed. Monoclonal antibodies 3E6 and ER-PR8 have been used to assess PSMA and PSA expression respectively. The evaluation of angiogenesis has been made by CD34 immune marker. Serum levels of PSA have been assayed by Immulite autoanalyser. RESULTS: The study of each protein separately among sera PSA levels showed that PSMA expression and angiogenic activity have the highest intensity in PC patients with serum PSA levels>20 ng/mL. Nevertheless, the lowest tissue PSA expression was found in PC patients with this latter sera PSA group. The most relevant results showed that in PC patients (PSA+, PSMA+) and (PSA-, PSMA+) profile were found to be inversely related to sera PSA levels. In PC patients, a high immunoexpression of (PSA+, PSMA+) profile has detected in the sera PSA group>20 ng/mL; whereas a high immunoexpression of (PSA-, PSMA+) profile was detected in the sera PSA group between 0 and 4 ng/mL. The highest angiogenic activity was found in PC patients with (PSA+, PSMA+) profile. CONCLUSIONS: Our findings clearly have supported the feasibility of PSA-PSMA profiles to improve in vivo diagnostic and therapeutic approaches in prostate cancer patients.

Impact of sampling origin on molecular detection of high-risk human papillomavirus and oncogene expression.

PURPOSE OF INVESTIGATION: The recognition of high-risk human papillomavirus (HR-HPV) as an etiological agent of cervical cancer has increased the importance of testing for HPV, and this might contribute to better risk stratification. METHODS: Eighty-eight randomly selected cervical cancer specimens including biopsies and their respective smears were used in this study. Control scrapings were obtained from ten healthy women. The presence of HPV16 and HPV18 was investigated using the technique of polymerase chain reaction (PCR) with the specific primers for the L1 region, while mRNA expression of HPV16 E6-E7 was evaluated by a reverse transcription PCR method (RT-PCR). RESULTS: The positivity for the viral genotype was influenced by the quantity of amplified DNA used. In tumor biopsies the higher positivity for HPV16 (54.5%) and HPV18 (15.9%) was obtained using 687.4 ng of DNA. At smears level solely 31.8% of HPV16 was detected using an average DNA quantity of about 157.2 ng. The revelation of HPV types depends on clinicopathologic data; HPV16 was detected more in advanced stages of squamous carcinoma (SC) samples (20% stage I, 62% Stage II and 80% stage III), while HPV18 and double infection were found exclusively at advanced stages of SC and in adenocarcinoma (AC), respectively (60%, 40% stage III SC and 80%, 20% Stage II A and C). The prevalence of HPV16 E6-E7 transcripts was evaluated at tumor biopsy with frequencies of 50%. CONCLUSION: Our data provide prospective evidence that HPV16/18L1 revelation at biopsy toward pathological types is efficient and correlates well with oncogenic transcript findings. Subtle changes in viral oncogene dynamics highlight the presence of other regulating proteins serving as additional biomarkers.

Profile of NF-κBp(65/NFκBp50) among prostate specific antigen sera levels in prostatic pathologies.

AIM: The aim of this work was to characterise the immunoexpression of NF-κB (p50/p65) in human prostatic pathologies and to study its profiles of activation among sera prostate specific antigen antigen (PSA) according the three groups: 0-4ng/mL, 4-20ng/mL and >20ng/mL. PATIENTS AND METHODS: Twenty-four men with benign prostate hyperplasia (BPH); 19 men with prostate cancer (PC) and five men with normal prostates (NP). Immunohistochemical and western blot analysis was performed. Serum levels of PSA were assayed by immulite autoanalyser. RESULTS: In BPH and PC samples, immunoexpressions were observed for NF-κBp65 and NF-κBp50; while in NP samples, only were detected NF-κBp50. PC samples showed immunoreactions to NF-κBp65 and NF-κBp50 more intense (respectively 24.18±0.67 and 28.23±2.01) than that observed in BPH samples (respectively18.46±2.04 and 18.66±1.59) with special localisation in the nucleus. Different profiles of NF-κBp65 immunoexpressions were observed and BPH patients with sera PSA levels between 0-4ng/mL presented a significant weak percentage compared to BPH patients with sera PSA levels between 4-20ng/mL and >20ng/mL. No immunoreactions to NF-κBp65 were observed in PC patients with sera PSA levels between 4-20ng/mL. CONCLUSION: The sensibility of both NF-κB and PSA to inflammation allowed confirming the relationship between these two molecules and its involvement in prostatic diseases progression (inflammatory and neoplasic).

[IL-2/IL-2R pathway in dendritic cell modulates both their cytokine synthesis profiles and their capacity to activate allogeneic CD4+ T lymphocytes]. / La voie IL-2/IL-2R chez la cellule dendritique module à la fois leur synthèse de cytokines et leur capacité à activer des lymphocytes T CD4+ allogéniques.

The results provide new insights into the role of IL-2/IL-2R pathway in DC. We report that stimulation of human monocyte-derived DC with LPS strongly upregulated CD25 (α chain of the IL-2R) expression. In addition, by using a humanized monoclonal antibody against CD25, we demonstrated that the IL-2 signalling in DC upregulated both IL-12 and γIFN production but decreased IL-10 synthesis. Anti-CD25 treatment reduced the ability of LPS-DC to induce allogeneic CD4(+) T cell proliferation as compared to LPS-matured DC. In addition, LPS-matured DC treated with IL-2 had a higher allostimulatory capacity compared to LPS-DC. We also found that LPS-matured DC produced IL-2. Thus, IL-2 seems to contribute actively to DC activation through an autocrine pathway. Moreover, IL-2 pathway in DC is involved in T helper priming. These findings might be useful for protocols in cellular therapy and a valuable tool to understand graft rejection versus the acquisition of peripheral tolerance.

Antibody response for L1, E6, E7 HPV 16, and HPV 18 antigens in Tunisian women with cervical cancer.

Results obtained in the present work indicated that the Luminex assay is more sensitive than ELISA. The reactivity to the early antigens E6 and E7 was 37% versus 42% for HPV 16 and 21% versus 20% for HPV 18 among cervical cancer cases using ELISA. However, these ratios were 44% and 61%, respectively, for E6 and E7 HPV 16 versus 28% and 21% for E6 and E7 HPV 18 when using the Luminex technique. Data also indicated that HPV 16 and HPV 18 showed distinct profiles for the different antigens tested. Finally, the differences in antibody responses between cervical cancer cases and benign cases toward the different antigens were significant.

Serological study in Tunisian cervical cancer patients.

OBJECTIVE: The aim of this study is to use a novel ELISA, based on five recombinant HPV-16 and HPV-18 proteins, for detection HPV-specific antibodies in a case-control study. PATIENTS AND METHODS: L1, E6 and E7 genes have been over expressed in Escherichia coli as double fused proteins. These recombinant proteins were used in a GST-capture ELISA as coating antigens. Human sera were collected from women with cervical cancer. Negative human sera were collected from patients apparently healthy and may be affected by other infectious agents. RESULTS: Most of the sera showed a positive reactivity to at least one of the HPV-16 or HPV-18 proteins (52/71). A percentage of 39.50% of the sera from HPV-16 infected women and 21.12% of the sera from women infected by HPV-18 genotype recognised at least one of the HPV-16 or HPV-18 proteins. Sera showed different reactivity to L1, E6 and E7 antigens, and only a few serum samples reacted to L1, E6 and E7 HPV-16, E6 and E7 HPV-18 (co-infection). Differences of reactivity between cases and controls were significant (P<0.0001). CONCLUSION: This novel ELISA, based on recombinant HPV-16 and HPV-18 antigens, is able to detect antibodies in women infected by HPV genotypes. The assay is easy to perform and has low cost, making it suitable for monitoring the natural history of HPV infections as well as for detecting pre-existing HPV antibodies in women who receive vaccination.

Antibody response for L1, E6 and E7 HPV 16 and HPV 18 antigens in Tunisian women with cervical cancer and controls.

Results obtained in the present work indicated that the Luminex assay is more sensitive than ELISA. The reactivity to the early antigens E6 and E7 was 37% versus 42% for HPV 16 and 21% versus 20% for HPV 18 among cervical cancer cases using ELISA. However, these ratios were 44% and 61%, respectively, for E6 and E7 HPV 16 versus 28% and 21% for E6 and E7 HPV 18 when using the Luminex technique. Data also indicated that HPV 16 and HPV 18 showed distinct profiles for the different antigens tested. Finally, the differences in antibody responses between cervical cancer cases and benign cases toward the different antigens were significant.

Anti-CD25 antibodies affect cytokine synthesis pattern of human dendritic cells and decrease their ability to prime allogeneic CD4+ T cells.

Anti-CD25 monoclonal antibodies are widely used in clinical transplantation to prevent acute allograft rejection. Although their effects on T lymphocytes have been extensively studied, their impact on human dendritic cells (DC) has never been reported. Furthermore, the role of the IL-2 in DC functions has not yet been fully elucidated. In this study, we confirm that the stimulation of human monocyte-derived DC with LPS strongly induced the expression of CD25 and that LPS-matured DC also expressed the beta and gamma chain of the IL-2R. We also showed that adding anti-CD25 monoclonal antibodies to LPS induced a decrease in IL-12, IL-1, TNF-alpha, IL-6, and IFN-gamma production and an increase in IL-10 synthesis by DC compared with stimulation with LPS alone. Furthermore, we showed that these modifications diminished the T helper priming ability of DC and polarized the alloimmune response toward TH2. In contrast, humanized anti-CD25 monoclonal antibodies did not affect the up-regulation of CD86, CD80, CD83, HLADR, or CD40 induced upon LPS stimulation. Taken together, this study discloses some previously unrecognized effects of anti-CD25 monoclonal antibodies on DC that may contribute to their clinical efficacy. In addition, this study also shed some light on the role of the IL-2 in human DC activation.

Formation and persistence of DNA adducts in epidermal and dermal mouse skin exposed to benzo(a)pyrene in vivo.

Tritiated benzo(a)pyrene was applied topically to 35 old male Swiss mice that received 250 nMole (0.63 mCi) per mouse in 150 microliters acetone. At each time point of study, 1, 3 and 7 days, 10 animals were killed, the skin was removed and the susceptible epidermic cells were separated from resistant dermis. The initial level of the total BaP-DNA adduct after 1 day of test was 2 fold higher in epidermic cells; in addition, the concentration of the individual modified deoxyribonucleoside adducts was 4 fold greater in epidermis. However, the nature and the repartition of the modified nucleosides analyzed by high performance liquid chromatography were similar. They were 5 fold more in epidermic cells, except for the (+/-) SynBaP- diolepoxide - deoxyguanosine adduct which was 2 fold more in dermic cells. The major DNA adduct formed in both types of cells was the (+) antiBaP diolepoxide - deoxyguanosine = (+) anti BPDE-dG with 75% of total adduct in epidermic and 55% in dermal cells. The decreased amount of BaP bound to DNA of epidermic and dermic cells may be similar and 90% of (+) anti BPDE-dG was removed after a week of treatment; in addition, a minority that bound with 9OH-BaP was also shown to be persistent. Although the persistence of adduct was 7 fold more important in susceptible epidermal cells than in the resistant dermic population, the nature of adduct repartition and the similar type of excision suggest that other mechanisms are also involved in the biologically different response of epidermis versus dermis to the carcinogenic BaP when it is applied topically.

Generation of lymphokine-activated killer cells by adherent LGL phenotype cells and non-adherent T lymphocytes.

In this work we separated human blood lymphocytes (PBL) in two populations (A-LAK and NA-LAK cells) by the adherence plastic method. A maximum adherence of cells was obtained after 2 days of PBL incubation in LAK medium containing 500 U/ml rIL-2. The A-LAK cells had LGL phenotype but 40% of them had a macrophage phenotype marker and less than 20% weakly expressed a T-cell marker. This population, when reincubated in culture, produced an increasing titre of interferon. At the same time, a significant NK activity against K562 target cells was measured just after enrichment; these enriched adherent cells also developed an increased LAK activity against DAUDI cell lines, ninefold more at 6 days than when assayed just after enrichment. In contrast, 75% of the NA-LAK enriched cells expressed T-cell marker; these produced two- to threefold less interferon than A-LAK cells at all time-points. The NA-LAK lymphocytes enhanced principally LAK activity measured by 70% lysis against DAUDI target cells tested at 6 days of culture. Further studies are in progress to determine the nature of the effector cells that mediated LAK activity.

High interferon titer and defective NK-cell activity in the circulation of nasopharyngeal carcinoma patients.

The interferon (IFN) activity of sera from 19 patients with nasopharyngeal carcinoma (NPC) was determined by the plaque-reduction assay with vesicular stomatitis virus (VSV) in HeLa cells and compared to that of sera from matched healthy controls. High titers of interferon were detected in the sera of the NPC patients with a geometric mean titer (GMT) of 43 +/- 25 U/ml. The interferon activity of the patients' sera was acid- and heat-labile (pH = 2 and 56 degrees C for 1 hr) and could be neutralized by a goat antiserum to human IFN-gamma. Interferon titers of the patients, in contrast, to normal controls, were not correlated with natural killer (NK) activity which was abnormally low in the NPC patients. On the other hand, a high percentage of circulating cells co-expressing the LGL marker (HNK-I) and the OKT8 antigen was detected in parallel with high IFN levels in NPC patients.

[Abdominal cystic lymphangioma. Apropos of a case in an adult]. / Lymphangiome kystique abdominal. A propos d'un cas chez l'adulte.

The authors report a case of cystic abdominal lymphangioma that appeared 4 months after delivery in a woman of 31 years of age. As in most of these cases, the exact diagnosis was not made before operation. Its occurrence following recent pregnancy makes one think that it could have arisen from a lymphatic block.