Cytokines are early diagnostic biomarkers of graft-versus-host disease in liver recipients

BACKGROUND： Graft-versus-host disease （GVHD） is associated with high mortality. Early diagnosis is essential to start treatment and to improve outcomes. Because of the inflammatory nature, we hypothesis that cytokine profile of patients with GVHD may serve as diagnostic markers. The present study was to evaluate the role of cytokine profile in the diagnosis of GVHD. METHODS： An immunoassay was used to detect 29 cytokines simultaneously in the serum; the measuring sensitivity of all cytokines was pg/mL. Healthy subjects undergoing annual routine physical examinations served as negative controls; 23 patients with hepatocellular carcinoma （HCC） who had undergone liver transplantation （the LT group） comprised the test subjects. A total of 22 kidney recipients with biopsyconfirmed GVHD （the RT group） were included for comparison. HCC patients with radical surgery （the HCC group, n=22） served as positive control. The liver contents of the three cytokines, IL-2, IL-18, and IFN-γ, were detected with immunohistochemistry. Serum granzyme B and perforin were measured by flow cytometry.RESULTS： Of the 29 cytokines, the levels of IL-2 and IL-18 were increased significantly in liver recipients with GVHD compared with healthy controls （P〈0.05）. The serum levels of these three cytokines in the healthy, HCC, LT, and RT groups were IL-2： 0.90±0.02, 4.14±0.61, 5.10±0.89, and 1.48±0.09 pg/mL; IL-18： 80.61±9.35, 109.51±10.93, 230.11±12.92, and 61.98±7.88 pg/mL; IFN-γ： 24.06±3.88, 24.84±3.21, 40.37±5.88, and 15.33±4.72 pg/mL, respectively. Immunohistochemistry showed that these 3 cytokines expressions in the liver were parallel to the serum cytokine. After standard anti-GVHD treatment, the expressions of IL-2, IL-18, and IFN-y were de- creased in the liver （P〈0.05）. Serum granzyme B and perforin were significantly increased in GVHD patients （P〈0.05）. CONCLUSIONS： IL-2, IL-18 and IFN-γ were from liver and might serve as biomarkers for monitoring GVHD develop- ment and the effects of anti-GVHD treatment. Granzyme B and perforin may play a role in increasing IL-2, IL-18, and IFN-y levels in GVHD patients.

Influence of HLA class Ⅱ molecules on the outcome of hepatitis B virus infection in population of Zhejiang Province in China

OBJECTIVE: To investigate the correlation between HLA class Ⅱ molecules and the different outcome of viral hepatitis B. METHODS: Thirty patients with chronic hepatitis B and 56 subjects who had spontaneously recovered from HBV infection in Zhejiang were enrolled in this investigation. HLA class Ⅱ molecules types and alleles were determined by PCR-ssp. RESULTS: HLA-DR12 was found in 21 of the 56 subjects (38%) recovered from hepatitis B, compared to 3 of the 30 patients with chronic hepatitis B [10%, relative risk (rr), O.19; P_(corr)<0.025]. The frequency of the allele of HLA-DR12 DRBl~* 1201 was higher in the subjects recovered from hepatitis B infection (32%) than in the patients with chronic hepatitis B (3%; rr, 0.07; P_(corr)<0.005). On the contrary, more HLA-DR9 was detected in the patients with chronic hepatitis B (43%) than in the subjects who recovered from hepatitis B infection (18%; rr, 3.52; P_(corr)<0.025). HLA-DQ9 was also detected with a higher frequency in the patients with chronic hepatitis B (43%) than in the subjects who recovered from hepatitis B (20%; rr, 3.13; P_(corr)<0.05). CONCLUSIONS: The HLA class Ⅱ molecule DR12 and its allele DRBl~* 1201 are associated with protection against chronic hepatitis B in Zhejiang Province, China. HLA-DR9 and DQ9 are associated with chronicity of HBV infection.

Potential value of autoantibodies as biomarkers of chronic graft-versus-host disease after allogeneic stem cell transplantation

We investigated the value of autoantibodies as biomarkers of chronic graft-versus-host disease(cGVHD)by analyzing the autoantibody profiles of 65 patients(34 cGVHD and 31 non-cGVHD)surviving longer than three months after allogeneic hematopoietic stem cell transplantation(allo-HSCT).Autoantibodies to at least one antigen were detected in 45 patients(70.8%),with multiple autoantibodies detected in 30 patients(46.2%).Antinuclear antibodies(ANAs)were the most frequently detected autoantibodies,with a significantly higher prevalence in non-cGVHD patients and c GVHD patients than that in healthy controls(HCs).ANA-nucleolar(ANA-N)was the main immunofluorescence pattern of ANA-positivity in both the non-cGVHD and c GVHD groups.There was a higher prevalence of anti-Ro52-positivity in non-cGVHD and cGVHD patients than in HC.Liver cGVHD was significantly associated with anti-Ro52-positivity.However,cGVHD activity and severity were not associated with the presence of autoantibodies.Similarly,there were no significant differences in overall survival or relapse among the four groups of patients expressing autoantibodies.Our results suggest that autoantibodies have limited value in predicting cGVHD.