Short-term ex vivo tissue culture models help study human lung infectionsA review.

Most studies on human lung infection have been performed using animal models, formalin or other fixed tissues, and in vitro cultures of established cell lines. However, the experimental data and results obtained from these studies may not completely represent the complicated molecular events that take place in intact human lung tissue in vivo. The newly developed ex vivo short-term tissue culture model can mimic the in vivo microenvironment of humans and allow investigations of different cell types that closely interact with each other in intact human lung tissues. Therefore, this kind of model may be a promising tool for future studies of different human lung infections, owing to its special advantages in providing more realistic events that occur in vivo. In this review, we have summarized the preliminary applications of this novel short-term ex vivo tissue culture model, with a particular emphasis on its applications in some common human lung infections.

Corrigendum: Expanded CURB-65: a new score system predicts severity of community-acquired pneumonia with superior efficiency.

This corrects the article DOI: 10.1038/srep22911.

Expanded CURB-65: a new score system predicts severity of community-acquired pneumonia with superior efficiency.

Aim of this study was to develop a new simpler and more effective severity score for community-acquired pneumonia (CAP) patients. A total of 1640 consecutive hospitalized CAP patients in Second Affiliated Hospital of Zhejiang University were included. The effectiveness of different pneumonia severity scores to predict mortality was compared, and the performance of the new score was validated on an external cohort of 1164 patients with pneumonia admitted to a teaching hospital in Italy. Using age ≥ 65 years, LDH > 230 u/L, albumin < 3.5 g/dL, platelet count < 100 × 10(9)/L, confusion, urea > 7 mmol/L, respiratory rate ≥ 30/min, low blood pressure, we assembled a new severity score named as expanded-CURB-65. The 30-day mortality and length of stay were increased along with increased risk score. The AUCs in the prediction of 30-day mortality in the main cohort were 0.826 (95% CI, 0.807-0.844), 0.801 (95% CI, 0.781-0.820), 0.756 (95% CI, 0.735-0.777), 0.793 (95% CI, 0.773-0.813) and 0.759 (95% CI, 0.737-0.779) for the expanded-CURB-65, PSI, CURB-65, SMART-COP and A-DROP, respectively. The performance of this bedside score was confirmed in CAP patients of the validation cohort although calibration was not successful in patients with health care-associated pneumonia (HCAP). The expanded CURB-65 is objective, simpler and more accurate scoring system for evaluation of CAP severity, and the predictive efficiency was better than other score systems.

Intrahepatic levels of PD-1/PD-L correlate with liver inflammation in chronic hepatitis B.

BACKGROUND AND OBJECTIVE: Programmed cell death-1 (PD-1) represents a mechanism of T-cell dysfunction in hepatitis B virus (HBV) persistence. In peripheral blood, PD-1 is up-regulated in virus-specific T cells, leading to the impairment of T cells. This study investigated the intrahepatic expression of PD-1 and its ligand (PD-L) in patients with chronic hepatitis B (CHB) virus. METHODS: Liver specimens were obtained from CHB (n = 56), acute hepatitis B (AHB, n = 12) patients and age-matched healthy subjects (n = 10). The expression of PD-1/PD-L was determined by immunohistochemistry. RESULTS: In CHB patients, PD-1 was predominantly expressed in lymphocytes infiltrating the portal tract. PD-L1 was detected in lymphocytes, hepatocytes and liver sinusoidal endothelial cells, while PD-L2 was localized in Kupffer cells and dendritic cells. The labeling indexes of PD-1 and PD-L1 in lymphocytes infiltrating portal area were significantly higher in CHB patients than in healthy controls and AHB patients. Within the CHB patients, the increases in labeling indexes of PD-1 and PD-L paralleled the degree of inflammation. CONCLUSIONS: These results suggest that over-expression of PD-1, PD-L1 and PD-L2 within liver may participate in local immune dysfunction, which could be one of the mechanisms involved in the chronicity of HBV infection and chronic inflammation seen in CHB patients.

Pegylated interferon α-2b up-regulates specific CD8+ T cells in patients with chronic hepatitis B.

AIM: to investigate the effect of pegylated interferon (IFN) α-2b on specific CD8+ T lymphocytes in patients with chronic hepatitis B (CHB). METHODS: twenty-one patients with CHB were treated with pegylated IFN α-2b. Periphery blood mononuclear cells were isolated from fresh heparinized blood by Ficoll-Hypaque density gradient centrifugation (density: 1.077 g/L, Pharmingen) at weeks 0, 4, 8, 12, and 24, respectively. Frequency of circulating hepatitis B virus (HBV) epitope-specific CD8 T cells was detected by flow cytometry. Cytokines were detected by cytometric bead assay. RESULTS: the frequency of circulating HBV core or env-specific CD8 T cells was higher (P < 0.05), the number of HBV core specific CD8 T cells was greater at week 24 (P < 0.05), the level of Th1-type cytokines [interleukin (IL)-12, tumor necrosis factor-α, and IFN-γ] was higher, while that of Th2-type cytokines (IL-4, IL-6, and IL-10) was lower in responders than in non-responders (P < 0.05) after pegylated IFN α-2b treatment. The IL-6 level was correlated with HBV DNA (r = 0.597, P = 0.04), while the inducible protein-10 (IP-10) level was correlated with serum alanine aminotransferase (ALT) (r = 0.545, P = 0.005). The IP-10 level at week 8 after pegylated IFN α-2b treatment could predict the normalization of ALT in CHB patients (positive predict value = 56%, negative predict value = 92%). CONCLUSION: pegylated IFN α-2b can enhance the immune response of CHB patients by increasing the frequency of HBV specific CD8+ T cells and regulating the Th1/Th2 cytokines.

Baseline predictors of virological response for chronic hepatitis B patients.

AIM: To determine which baseline factors of chronic hepatitis B patients are predictive of virological response to Peginterferon alpha-2b therapy. METHODS: A total of 21 HBeAg-positive chronic hepatitis B (CHB) patients treated with Peginterferon alpha-2b were recruited. They were treated with Peginterferon alpha-2b (0.5-1.0 microg/kg per week) for 24 wk and followed up for 24 wk. Clinical and laboratory data of the patients were determined at pretreatment and at week 12, at 24 during treatment, and at week 48 during follow up. RESULTS: Ten patients achieved a virological response at the end of treatment. Their baseline serum alanine aminotransferase (ALT), thyroid-stimulating hormone (TSH), and total thyroxin (TT4) levels were significantly different from those who failed treatment. The positive predictive values (PPV) and negative predictive values (NPV) of ALT, TSH, and TT4 were 75% and 89 %, 75% and 89 %, and 75% and 75%, respectively. Moreover, combinations of the baseline ALT and TT4, ALT and TSH, TT4 and TSH levels had much higher PPV and NPV (86% and 88%, 89% and 100%, 83% and 100%, respectively). CONCLUSION: Baseline serum ALT, TSH, and TT4 levels, especially in combination, have high predictive values of virological response to Peginterferon alpha-2b in HBeAg-positive CHB patients.

Hepatoma cells up-regulate expression of programmed cell death-1 on T cells.

AIM: To investigate the effect of hepatoma cells on up-regulation of programmed cell death-1 (PD-1), and the function of PD-1 on T cells. METHODS: HepG2 or HepG2.2.1.5 cells were co-cultured with a lymphoma cell line-Jurkat cells. PD-1 expression was detected by flow cytometry. IL-2, INF-gamma and IL-10 in culture supernatant were detected by enzyme-linked immunosorbent assay (ELISA). Cytotoxic action of T cells was determined by MTT reduction assay-direct mononuclear cell cytotoxicity assay. RESULTS: The PD-1 expression on Jurkat cells increased by 16.17% +/- 2.5% and 17.43% +/- 2.2% after HepG2 or HepG2.2.1.5 cells were co-cultured for 48 h. The levels of IL-2, INF-gamma and IL-10 in the culture supernatant were 202.9 +/- 53.0 pg/mL, 88.6 +/- 4.6 pg/mL and 63.7 +/- 13.4 pg/mL respectively, which were significantly higher than those (102.9 +/- 53 pg/mL, 39.3 +/- 4.2 pg/mL, and 34.6 +/-13.7 pg/mL) in the control group (P < 0.05). The OD value for MTT assay in the blocking group (0.29 +/- 0.06) was significantly higher than that (0.19 +/- 0.09) in the control group (P < 0.05). CONCLUSION: PD-1 expression on Jurkat cells is up-regulated by hepatoma cells, cytokines and cytotoxic action are elevated after PD-1/PD-L1 is blocked.

[A clinical analysis of benign prostatic hyperplasia with chronic prostatitis].

OBJECTIVE: To study the incidence of benign prostatic hyperplasia (BPH) complicated by chronic prostatitis. METHODS: We performed routine examinations and bacterial culture of the expressed prostate secretion (EPS) for 213 cases of BPH, detected mycoplasma, chlamydia and serum PSA, and compared the results of IPSS of those complicated with chronic prostatitis before and after a 4-week anti-inflammatory treatment. RESULTS: Of the total cases, 69 (32.4%) were complicated by chronic prostatitis, 27 (12.7%) EPS positive and 15 (7.0%) mycoplasma and chlamydia positive. Among the 69 cases of chronic prostatitis, 7 were found with an elevated level of PSA (> 4 microg/L), and 43 with the mean IPSS score decreased from (12.2 +/- 2.6) before anti-inflammatory treatment to (10.5 +/- 2.3) after it (P < 0.01). CONCLUSION: EPS examination should be performed for patients with BPH, which is highly significant for the diagnosis of prostatitis, choice of medical or surgical treatment, improvement of therapeutic effect and reduction of complications.

[HBV transinfected enhances the PD-L expression of cytokines stimulating].

OBJECTIVE: To investigate whether the PD-L expression in the liver cell lines transinfected with HBV (HepG2.2.15 cells) can be up-regulated after cytokines stimulating. METHODS: To apply the liver cell lines (HepG2 cells and HepG2.2.15 cells) as a model, the cells were stimulated with IL-4, IFN-alpha and IFN-gamma (final concentration were 10 ng/ml, stimulated for 12 hours) and RT-PCR was carried out to determine the PD-L expression before and after cytokines stimulating. RESULTS: Whether or not transinfected with HBV, IFN-alpha and IFN-gamma both can induce the liver cell lines (HepG2 cells and HepG2.2.15 cells) PD-L1 expression while IL-4 can not; IL-4, IFN-alpha, IFN-gamma all can induce the PD-L2 expression in HepG2.2.15 cells which was transinfected with HBV, only IFN-gamma can induce the PD-L2 expression in HepG2 cells which was not transinfected with HBV. CONCLUSION: IFN-alpha, IFN-gamma both can induce the PD-L1 expression in HepG2 cells and HepG2.2.15 cells, while it is easy for cytokines to induce the PD-L2 expression in HepG2.2.15 cells which was transinfected with HBV, this may provide a potential mechanism of the molecular basis for chronic HBV infection.