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Objective To explore the effect of the precise diagnosis and treatment of a case of acute graft-versus-host disease （aGVHD） with rash as the main manifestation under the guidance of immune monitoring. Methods Clinical data and diagnosis and treatment of a child with high-risk acute myeloid leukemia who developed intractable rash after receiving unrelated umbilical cord blood transplantation were retrospectively analyzed. Using the keywords of “hematopoietic stem cell transplantation”“immune reconstitution” and “acute graft-versus-host disease”， relevant literatures were searched from PubMed， Web of Science， CNKI and Wanfang databases， and relevant cases were collected and analyzed. Results Hematopoietic reconstruction was successfully achieved in this 1 year 9 months old girl. On day 22 after transplantation， the skin of head and back neck of the patient showed dense red papules （approximately 19% in area）， with obvious itching. Under the treatment of oral tacrolimus and topical hormone drugs， the rash was not alleviated （area>90%）. Meanwhile， the proportion of CD3+ cells， CD8+ cells， CD3+CD69+cells and CD3+HLA-DR+cells was significantly increased， and the proportion of Treg cells was decreased. The immune indexes supported T lymphocyte activation， then diagnosis of gradeⅡaGVHD （skin grade 3） was made， and the immunosuppressive regimen was strengthened. During this period， the skin rash of the child subsided， but recurred， and accompanied by obvious skin desquamation. The proportion of CD3+ cells， CD8+ cells， and CD3+HLA-DR+cells remains relatively high， supporting the diagnosis of aGVHD. Therefore， the anti-GVHD regimen was adjusted and strengthened， and the aGVHD of the child’s skin was mitigated. Regular monitoring showed that the primary disease was completely relieved， the implantation rate was 100%， and the immune reconstitution was stable. During 32-month follow-up after transplantation， the child obtained disease-free survival. Nine relevant literatures were retrieved， all of which demonstrated that the increased proportion of early CD3+ cells， CD8+ cells， activated T lymphocytes and Treg downregulation were associated with the occurrence of aGVHD. Conclusions The dynamic monitoring of the immune status of patients after allogeneic hematopoietic stem cell transplantation is helpful to evaluate their immune reconstitution and disease status （such as， aGVHD）， thereby assisting clinicians to formulate a reasonable immunosuppressive regimen and bring favorable prognosis to patients.

Objective To investigate the long-term safety and effectiveness of withdrawal of hepatitis B immuneglobulin (HBIG) and/or nucleos(t)ide analogues (NAs) to prevent hepatitis B virus (HBV) reinfection in liver transplant recipients with hepatitis B-related diseases after successful vaccination. Methods Baseline data of 76 liver transplant recipients undergoing hepatitis B immune reconstitution after receiving hepatitis B vaccines were retrospectively analyzed. The vaccination and response, the follow-up results of respondents with HBIG and/or NAs withdrawal, and the reinfection of HBV after withdrawal of HBIG and/or NAs were analyzed. Results The time interval from liver transplantation to hepatitis B vaccination was 26 (20, 40) months. The time interval from vaccination to response was 15 (8,27) months. Initially, 76 recipients withdrew HBIG, and 36 recipients withdrew HBIG and NAs. During the follow-up, 12 of 76 recipients who withdrew HBIG resumed use of HBIG, and 16 of 36 recipients who withdrew HBIG and NAs resumed use of NAs. The withdrawal time of HBIG and NAs was 135 (98,150) and 133 (34,149) months, respectively. Sixteen respondents did not receive booster, and 36 respondents received boosters on a regular basis. The time interval between the first booster and HBIG withdrawal was 44 (11,87) months. No significant differences were observed in baseline data between the respondents with and without boosters (all P>0.05). During the follow-up, 9 recipients were lost to follow-up, 5 were re-infected with HBV, 3 died, and 1 recipient developed graft loss and underwent secondary liver transplantation. Among 5 recipients re-infected with HBV, 4 cases had virus mutation. Significant differences were found between re-infected and uninfected patients regarding withdrawal of NAs and hepatitis B e antigen (HBeAg) positive before transplantation (both P<0.05). Conclusions Long-term withdrawal of HBIG is feasible and safe for recipients with successful hepatitis B immune reconstitution after liver transplantation for hepatitis B-related diseases. Nevertheless, whether antiviral drugs can be simultaneously withdrawn remains to be validated. [ABSTRACT FROM AUTHOR]

Objective To explore the effect of the precise diagnosis and treatment of a case of acute graft-versus-host disease (aGVHD) with rash as the main manifestation under the guidance of immune monitoring. Methods Clinical data and diagnosis and treatment of a child with high-risk acute myeloid leukemia who developed intractable rash after receiving unrelated umbilical cord blood transplantation were retrospectively analyzed. Using the keywords of “hematopoietic stem cell transplantation”“immune reconstitution” and “acute graft-versus-host disease”, relevant literatures were searched from PubMed, Web of Science, CNKI and Wanfang databases, and relevant cases were collected and analyzed. Results Hematopoietic reconstruction was successfully achieved in this 1 year 9 months old girl. On day 22 after transplantation, the skin of head and back neck of the patient showed dense red papules (approximately 19% in area), with obvious itching. Under the treatment of oral tacrolimus and topical hormone drugs, the rash was not alleviated (area>90%). Meanwhile, the proportion of CD3+ cells, CD8+ cells, CD3+ CD69+ cells and CD3+ HLA-DR+ cells was significantly increased, and the proportion of Treg cells was decreased. The immune indexes supported T lymphocyte activation, then diagnosis of grade Ⅱ aGVHD (skin grade 3) was made, and the immunosuppressive regimen was strengthened. During this period, the skin rash of the child subsided, but recurred, and accompanied by obvious skin desquamation. The proportion of CD3+ cells, CD8+ cells, and CD3+ HLA-DR+ cells remains relatively high, supporting the diagnosis of aGVHD. Therefore, the anti-GVHD regimen was adjusted and strengthened, and the aGVHD of the child’s skin was mitigated. Regular monitoring showed that the primary disease was completely relieved, the implantation rate was 100%, and the immune reconstitution was stable. During 32-month follow-up after transplantation, the child obtained disease-free survival. Nine relevant literatures were retrieved, all of which demonstrated that the increased proportion of early CD3+ cells, CD8+ cells, activated T lymphocytes and Treg downregulation were associated with the occurrence of aGVHD. Conclusion The dynamic monitoring of the immune status of patients after allogeneic hematopoietic stem cell transplantation is helpful to evaluate their immune reconstitution and disease status (such as, aGVHD), thereby assisting clinicians to formulate a reasonable immunosuppressive regimen and bring favorable prognosis to patients. [ABSTRACT FROM AUTHOR]

Objective To observe the effect of Tonifying deficiency and Zhengqi porridge on immune function reconstruction of HIV/AIDS patients with Qi deficiency. Methods Sixty (Human Immunodeficiency Virus/ Acquired Immunodeficiency Syndrome)HIV/AIDS patients admitted to the center for Disease Control and prevention in Huichuan District of Zunyi City were randomly divided into control group and treatment group by random number table. All patients received the highly active anti-retroviral therapy, and those in the treatment group were given Tonifying deficiency and Zhengqi porridge. The CD4+ T-cell count was measured before, 3 months and 6 months after intervention. The Effective reconstitution of immune function was analyzed. Results After 3 and 6 months of treatment, the immune reconstitution in the treatment group was better than that in the control group for patients with CD4+ T-cell count of 100-500/mm3 (P＜0. 05). Conclusion Tonifying deficiency and Zhengqi porridge can promote the reconstruction of immune function in HIV / AIDS patients with Qi deficiency and help to restore immune function. (目的 探讨补虚正气粥对人类免疫缺陷病毒(HIV)感染及艾滋病(AIDS)气虚型患者的免疫功能恢复作用。方法 选择遵义市汇川区疾病预防控制中心60名HIV/AIDS患者, 随机将患者分为治疗组和对照组, 各30例。对照组按传统方法进行HAART(高效抗反转录病毒)治疗, 治疗组则在对照组治疗的基础上加用补虚正气粥。于干预前、干预后3个月和干预后6个月比较两组患者CD4+T淋巴细胞计数值和免疫功能重建有效率。结果 对于干预前CD4+T淋巴细胞计数100～500个/mm3的患者, 在介入3个月和6个月, 治疗组免疫功能的恢复较对照组好(P＜0. 05)。结论 补虚正气粥对HIV/AIDS气虚型患者免疫功能重建有促进作用, 有助于免疫功能的重建。)

Objective: To analyze immune reconstitution and influencing factors in HIV infected men who have sex with men (MSM) with access to antiviral therapy (ART) in Guangxi Zhuang Autonomous Region (Guangxi) during 2005-2021. Methods: The data were collected from Chinese Disease Prevention and Control Information System. The study subjects were HIV infected MSM with access to the initial ART for ≥24 weeks in Guangxi from 2005 to 2021 and HIV RNA lower than the detection limit within 24 months. The proportion of infected MSM who had immune reconstitution after ART was calculated. Cox proportional hazard regression model was used to analyze the influencing factors of immune reconstitution. Software SPSS 24.0 was used for statistical analysis. Results: A total of 3 200 HIV infected MSM were enrolled, in whom 15.56 % (498/3 200) had no immune reconstitution, 14.78% (473/3 200) had moderate immune reconstitution, and the rate of complete immune reconstitution was 69.66% (2 229/3 200). The M ( Q 1 ) of ART time for immune reconstitution was 12 (5, 27) months. Multivariate Cox proportional risk regression model analysis results showed that compared with those with initial ART at age ≥30 years, WHO clinical stage Ⅲ/Ⅳ illness, baseline BMI <18.50 kg/m Q 3 ) of ART time for immune reconstitution was 12 (5, 27) months. Multivariate Cox proportional risk regression model analysis results showed that compared with those with initial ART at age ≥30 years, WHO clinical stage Ⅲ/Ⅳ illness, baseline BMI <18.50 kg/m 2 and baseline CD4 counts ≥200 cells/µl were more likely to have complete immune reconstitution. + In the HIV infected MSM in Guangxi, failures to achieve moderate and complete immune reconstitution were observed. Surveillance and ART regimen should be improved for key populations, such as those with older age and low baseline CD4 counts.2 and baseline CD4 counts ≥200 cells/µl were more likely to have complete immune reconstitution. Conclusions: In the HIV infected MSM in Guangxi, failures to achieve moderate and complete immune reconstitution were observed. Surveillance and ART regimen should be improved for key populations, such as those with older age and low baseline CD4 counts.

Objective: To analyze the immune reconstitution after BTKi treatment in patients with chronic lymphocytic leukemia (CLL)., Methods: The clinical and laboratorial data of 59 CLL patients admitted from January 2017 to March 2022 in Fujian Medical University Union Hospital were collected and analyzed retrospectively., Results: The median age of 59 CLL patients was 60.5(36-78). After one year of BTKi treatment, the CLL clones (CD5 + /CD19 + ) of 51 cases (86.4%) were significantly reduced, in which the number of cloned-B cells decreased significantly from (46±6.1)×10 9 /L to (2.3±0.4)×10 9 /L ( P =0.0013). But there was no significant change in the number of non-cloned B cells (CD19 + minus CD5 + /CD19 + ). After BTKi treatment, IgA increased significantly from (0.75±0.09)g/L to (1.31±0.1)g/L ( P <0.001), while IgG and IgM decreased from (8.1±0.2)g/L and (0.52±0.6)g/L to (7.1±0.1)g/L and (0.47±0.1)g/L, respectively ( P <0.001, P =0.002). BTKi treatment resulted in a significant change in T cell subpopulation of CLL patients, which manifested as both a decrease in total number of T cells from (2.1±0.1)×10 9 /L to (1.6±0.4)×10 9 /L and NK/T cells from (0.11±0.1)×10 9 /L to (0.07±0.01)×10 9 /L ( P =0.042, P =0.038), both an increase in number of CD4 + cells from (0.15±6.1)×10 9 /L to (0.19±0.4)×10 9 /L and CD8 + cells from (0.27±0.01)×10 9 /L to (0.41±0.08)×10 9 /L (both P <0.001). BTKi treatment also up-regulated the expression of interleukin (IL)-2 while down-regulated IL-4 and interferon (IFN)-γ. However, the expression of IL-6, IL-10, and tumor necrosis factor (TNF)-α did not change significantly. BTKi treatment could also restored the diversity of TCR and BCR in CLL patients, especially obviously in those patients with complete remission (CR) than those with partial remission (PR). Before and after BTKi treatment, Shannon index of TCR in patients with CR was 0.02±0.008 and 0.14±0.001 ( P <0.001), while in patients with PR was 0.01±0.03 and 0.05±0.02 ( P >0.05), respectively. Shannon index of BCR in patients with CR was 0.19±0.003 and 0.33±0.15 ( P <0.001), while in patients with PR was 0.15±0.009 and 0.23±0.18 ( P <0.05), respectively., Conclusions: BTKi treatment can shrink the clone size in CLL patients, promote the expression of IgA, increase the number of functional T cells, and regulate the secretion of cytokines such as IL-2, IL-4, and IFN-γ. BTKi also promote the recovery of diversity of TCR and BCR. BTKi treatment contributes to the reconstitution of immune function in CLL patients.

Objective: To investigate the risk factors for human cytomegalovirus infection after allogeneic hematopoietic stem cell transplantation in children and the impact of human cytomegalovirus infection on post-transplant immune reconstitution. Methods: A Retrospective Co-Hort study design was used to include 81 children treated with allo-HSCT from January 2020 to March 2022 at the Department of Hematology, Capital Institute of Pediatrics, Beijing, China, and followed up for 1 year. Real-time quantitative PCR was used to detect positive detection of HCMV in children after allo-HSCT, multifactorial logistic regression modeling was used to analyze the risk factors leading to HCMV infection, and generalized estimating equation modeling was used to analyze the effect of HCMV infection on the T-cells of the children who received allo-HSCT. Results: The age M ( Q 1 , Q 3 ) of 81 children was 5.1 years (10 months, 13.8 years), and 50 (61.7%) were male. By the endpoint of follow-up, a total of 50 HCMV-positive cases were detected, with an HCMV detection rate of 61.7%; The results of multifactorial logistic regression modeling showed that children with grade 2-4 aGVHD had a higher risk of HCMV infection compared with grade 0-1 after transplantation [ OR (95% CI ) value: 2.735 (1.027-7.286)]. The results of generalized estimating equation modeling analysis showed that the number of CD3 + T cells in HCMV-positive children after transplantation was higher than that in the HCMV-negative group [ RR (95% CI ) value: 1.34 (1.008-1.795)]; the ratio of CD4 + T/CD8 + T cells was smaller than that in the HCMV-negative group [ RR (95% CI ) value: 0.377 (0.202-0.704)]; the number of CD8 + T cells was higher than that in the HCMV-negative group [RR (95% CI ) value: 1.435 (1.025-2.061)]; the number of effector memory CD8 + T cells was higher than that in the HCMV-negative group [ RR (95% CI ) value: 1.877 (1.089-3.236)]. Conclusion: Acute graft-versus-host disease may be a risk factor for HCMV infection in children after allo-HSCT; post-transplant HCMV infection promotes proliferation of memory CD8+T-cell populations and affects immune cell reconstitution.

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Objective: To observe the differences in biochemical indexes and AIDS-related complications at baseline in HIV-infected patients with different levels of immune reconstitution to antiretroviral therapy (ART). Methods: The subjects were treat-naïve adult HIV-infected patients who were followed up for more than 24 months in the Guangzhou Eighth People's Hospital affiliated infection clinic at Guangzhou Medical University from January 2010 to December 2017. CD4 + T lymphocyte count at baseline at <200, 200-350, and >350 cells/µl levels were divided into poor, partial, and good immune reconstitution groups. The Kruskal-Wallis H and chi-square tests were used to analyze the differences in baseline sociodemographic characteristics, biochemical indexes, and AIDS-related complications among different groups. The SPSS 20.0 software was used for statistical analysis. Results: Among the 3 900 HIV-infected individuals, 385 cases (9.9%), 1 206 cases (30.9%), and 2 309 cases (59.2%) were grouped into poor, partial and good immune reconstitution groups, respectively. The baseline biochemical indexes of leukocyte, platelet, hemoglobin, TG, TC, FPG, AST, ALT and total bilirubin in the poor immune reconstitution group were significantly different from those in the good immune reconstitution group (all P <0.05). The proportion of AIDS-related complications at baseline in the poor immune reconstitution group, such as tuberculosis, pneumocystis yeli pneumonia, disseminated mycosis, esophageal candidiasis, extrapulmonary tuberculosis, dermatitis, oral candidiasis, oral mucous leukoplakia, continuous diarrhea for more than 1 month and continuous or intermittent fever for more than 1 month, was significantly higher than that in the good immune reconstitution group (all P <0.05). Conclusions: The biochemical indexes and AIDS-related complications in HIV-infected patients with different levels of immune reconstitution were significantly different at baseline. Attention should be paid to monitoring abnormal biomedical indicators and AIDS-related complications at baseline.

Objective: To investigate the recovery characteristics of T cell subsets in patients with severe aplastic anemia (SAA) who received haploid hematopoietic stem cell transplantation(HSCT) and its relationship with acute graft-versus-host disease(aGVHD)., Methods: The clinical data of 29 SAA patients who received haploid hematopoietic stem cell transplantation in the department of hematology, Shanxi Bethune Hospital from June 2018 to January 2022 were retrospectively analyzed. The absolute counts of CD3 + T, CD4 + T, CD8 + T lymphocytes and the ratio of CD4 + T/CD8 + T lymphocytes in all patients before transplantation, 14, 21, 30, 60, 90 and 120 days after transplantation were analyzed. The proportion of T lymphocytes was compared in the non-aGVHD group, the grade Ⅰ-Ⅱ aGVHD group and the grade III-IV aGVHD group., Results: The counts of all T cells in 27 patients were far below the normal level at 14 and 21 days after transplantation, but there was obvious heterogeneity. There was a certain relationship between T cell immune reconstitution and conditioning regimen, age, and immunosuppressive treatment before transplantation. CD3 + T cells showed a steady upward trend at 30, 60, 90, and 120 days after transplantation, and returned to the normal levels at 120 days after transplantation; faster recovery of CD4 + T cells was closely related to aGVHD, which was at 30, 60, 90, 120 days after transplantation showed a slow upward trend, and which was still far below the normal level of 120 days after transplantation. CD8 + T cell counts began to recover at 14 and 21 days after transplantation, and the recovery was earlier than the CD4 + T cells, and its recovery speed was rapid 30 and 60 days after transptantation, which showed an upward trend and exceeded the normal levels 90 days after transplantation. Since CD8 + T cells reconstituted quickly, while the CD4 + T cells reconstitution was slowly, which made the long-term CD4 + T/CD8 + T cell ratio after transplantation was inverted . Compared with the non-aGVHD group, the absolute counts of CD3 + T, CD4 + T, and CD8 + T cells in the aGVHD group were significantly higher than those in the non-aGVHD group at each time period after transplantation. In the aGVHD group, grade Ⅲ-Ⅳ aGVHD occurred more frequently in the early post-transplantation period (within 14-21 days), the grade Ⅰ-Ⅱ aGVHD group mostly occurred within 30-90 days after transplantation, and CD3 + T, CD4 + T, CD8 + T cell counts in the grade Ⅲ-Ⅳ aGVHD group were significantly higher than those in the grade Ⅰ-Ⅱ aGVHD group; and the greater the proportion of CD4 + T, the more severe the degree of aGVHD., Conclusion: The speed of T cell immune reconstitution after SAA haploid transplantation is different, which is related to the conditioning regimen, age, and immunosuppressive therapy before transplantation. The rapid recovery of CD4 + T cells is closely related to the occurrence of aGVHD.

Objective: To investigate the clinical value of oligoclonal bands (OB) in patients with multiple myeloma (MM)., Methods: The laboratory test and clinical data of 624 newly diagnosed MM patients admitted to Blood Diseases Hospital of Chinese Academy of Medical Sciences from January 2013 to December 2019 were retrospectively analyzed, including 30 patients with OB, and the clinical characteristics, treatment effects and survival of OB and non-OB patients were analyzed and compared., Results: OB occurred in 11.8% (22/187) of patients who received autologous stem cell transplantation(ASCT) and only 1.8% (8/437) of patients who did not receive ASCT (P=0.000). The median time to the appearance of oligoclonal bands was 3.2(0.6-10.5) months after transplantation. The M protein types of oligoclonal bands mainly include IgG κ, IgG λ, IgM λ and λ light chains. In the presence of oligoclonal bands, 90% of patients were evaluated as complete remission (CR) and above. There were no statistically significant differences in disease stage, tumor burden, and genetic abnormalities between OB and non-OB patients. Among the all patients, the prognosis of OB patients was significantly better than that of non-OB patients, and OB patients showed deeper disease remission (significantly higher CR rate, MRD negative rate, and longer MRD negative duration). Among patients who underwent ASCT, OB patients showed earlier immune recovery, but the depth of treatment response and survival outcomes were similar between OB and non-OB patients, it was no statistically difference. Although OB patients showed earlier immune reconstitution, this did not translate into better survival, suggesting that the better prognosis of OB patients was mainly related to deeper and durable remission rather than early immune reconstitution. Further analysis in patients who received ASCT and obtained MRD negative indicated that there was no additional survival benefit in patients with OB., Conclusion: The better prognosis of OB patients may be related to the deeper treatment response, but not to the early immune reconstitution. The appearance of OB is only a sign of deep remission and early immune reconstitution in patients, it cannot be translated into survival benefit of MM patients.

Objective: To explore the relationship between the reconstitution of immune cells in patients with hematological malignancies and the occurrence of chronic graft-versus-host disease (cGVHD) after treatment with unrelated cord blood transplantation (UCBT) and sibling peripheral blood stem cell transplantation (PBSCT) . Methods: A total of 124 patients undergoing allogenic hematopoietic stem cell transplantation (allo-HSCT) in the First Affiliated Hospital of University of Science and Technology of China from March 2018 to August 2019, including 96 patients with UCBT and 28 patients with PBSCT. Peripheral blood immune cells of patients with UCBT and PBSCT were detected at 1, 3, 6, 9, and 12 months after transplantation using flow cytometry, and both UCBT and PBSCT patients were divided into cGVHD and non-cGVHD groups based on whether cGVHD occurred to explore the correlation between the immune cells reconstitution of the two types of transplantation and cGVHD. Results: ①The cumulative incidence of the moderate to severe cGVHD in the UCBT group was significantly lower than that in the PBSCT group[9.38% (95% CI 3.35%-15.02%) vs 28.57% (95% CI 9.72%-43.50%) , P =0.008]; the 2-year cumulative incidence of cGVHD and moderate to severe cGVHD in the UCBT group was lower than that in the PBSCT group[15.60% (95% CI 9.20%-23.60%) vs 32.10% (95% CI 15.80%-49.70%) , P =0.047; 10.40% (95% CI 5.30%-17.50%) vs 28.60% (95% CI 13.30%-46.00%) , P =0.014]. ②The absolute counts of CD4(+)T cells in the UCBT group were higher than those in the PBSCT group at 6, 9, and 12 months after transplantation[59.00 (36.70-89.65) ×10(7)/L vs 31.40 (18.10-44.00) ×10(7)/L, P <0.001; 71.30 (49.60-101.45) ×10(7)/L vs 41.60 (25.82-56.27) ×10(7)/L, P <0.001; 83.00 (50.17-121.55) ×10(7)/L vs 44.85 (31.62-62.10) ×10(7)/L, P <0.001]; the proportions of CD4(+)T cells in the UCBT group were always higher than those in the PBSCT group ( P <0.05) . The absolute counts and proportions of B cells in the PBSCT group were higher than those in the UCBT group at the first month after transplantation[0.70 (0.30-1.70) ×10(7)/L vs 0.10 (0-0.30) ×10(7)/L, P <0.001; 0.45% (0.30%-2.20%) vs 0.20% (0.10%-0.40%) , P =0.002]; the absolute counts and proportions of B cells in the UCBT group were higher than those in the PBSCT group at 9 and 12 months after transplantation[53.80 (28.00-103.20) ×10(7)/L vs 23.35 (5.07-35.00) ×10(7)/L, P <0.001; 21.45 (11.80-30.45) % vs 9.00% (3.08%-16.73%) , P <0.001. 66.70 (36.97-98.72) ×10(7)/L vs 20.85 (7.72-39.40) ×10(7)/L, P <0.001; 22.20% (14.93%-29.68%) vs 8.75% (5.80%-18.93%) , P <0.001]. The absolute counts and proportions of regulatory B (Breg) cells in the UCBT group were higher than those in the PBSCT group at 6, 9, and 12 months after transplantation[1.23 (0.38-3.52) ×10(7)/L vs 0.05 (0-0.84) ×10(7)/L, P <0.001; 5.35% (1.90%-12.20%) vs 1.45% (0-7.78%) , P =0.002. 2.25 (1.07-6.71) ×10(7)/L vs 0.12 (0-0.77) ×10(7)/L, P <0.001; 6.25% (2.00%-12.33%) vs 0.80% (0-5.25%) , P <0.001. 3.69 (0.83-8.66) ×10(7)/L vs 0.46 (0-0.93) ×10(7)/L, P <0.001; 6.15% (1.63%-11.75%) vs 1.40% (0.18%-5.85%) , P <0.001].The absolute counts and proportions of CD3(+)T cells, CD8(+)T cells, and Treg cells in the UCBT group were not significantly different from those in the PBSCT group. ③The absolute counts of B cells in the non-cGVHD group of UCBT patients were higher than those in the moderate to severe cGVHD group at 6 and 12 months after transplantation ( P =0.038, P =0.043) ; the proportions of B cells in the non-cGVHD group were higher than those in the moderate to severe cGVHD group at 6 months after transplantation ( P =0.049) . The absolute counts of Breg cells in the non-cGVHD group of patients with UCBT were higher than those in the moderate to severe cGVHD group at 6, 9, and 12 months after transplantation ( P =0.006, P =0.028, P =0.050) ; the proportions of Breg cells in the non-cGVHD group were higher than those in the moderate to severe cGVHD group at 9 months after transplantation ( P =0.038) . ④The absolute counts and proportions of B and Breg cells in the non-cGVHD group of patients with PBSCT were not statistically different than those in the moderate to severe cGVHD group. Conclusion: In the process of immune cell reconstitution, the Breg cells in the UCBT group were higher than those in the PBSCT group, and the Breg cells in the non-cGVHD group of the two types of transplantation were always higher than those in the moderate to severe cGVHD group, indicating that Breg cells can reduce the occurrence of cGVHD, revealing the possible reason for the lower incidence of cGVHD in the UCBT group.

Objective: To investigate whether interleukin-12 (IL-12) over-expression in malignant melanoma B16 cells affects the expression level of programmed death-1 (PD-1) on T cells in mice during immune microenvironment reconstruction., Methods: B16 cells were transfected with an IL-12 expression lentiviral vector, and IL-12 over-expression in the cells was verified qPCR and ELISA. Plate cloning assay was used to compare the cell proliferation activity between B16 cells and B16/IL-12 cells. The expression of IL-12 protein in B16/IL-12 cells-derived tumor tissue were detected by ELISA. C57BL/6 mice were inoculated with B16 cells or B16/IL-12 cells, and 14 days later the proportion of T cells with high expression of PD-1 in the tumor-draining lymph nodes was detected by flow cytometry. Mouse models of immune reconstitution established by 650 cGy X-ray radiation were inoculated with B16 (B16+RT group) or B16/IL-12 (B16/IL-12+RT group) cells, with the mice without X-ray radiation prior to B16 cell inoculation as controls. Tumor growth in the mice was recorded at different time points, and on day 14, flow cytometry was performed to detect the proportion of T cells with high PD-1 expression in the tumor-draining lymph nodes and in the tumor tissue., Results: B16 cells infected with the IL-12-overexpressing lentiviral vector showed significantly increased mRNA and protein levels of IL-12 ( P &lt; 0.001) without obvious changes in cell viability ( P &gt;0.05). B16/IL-12 cells expressed higher levels of IL-12 than B16 cells in vivo ( P &lt; 0.01). In the tumor-bearing mouse models, the proportion of CD4 + PD-1 + T cells was significantly lower in B16/IL-12 group than in B16 group ( P &lt; 0.01). In the mice with X-ray radiation-induced immune reconstitution, PD-1 expressions on CD4 + T cells ( P &lt; 0.05) and CD8+ T cells ( P &lt; 0.01) were significantly higher in B16+ RT group than in the control mice and in B16/IL-12+RT group ( P &lt; 0.01 or 0.001); the tumors grew more slowly in B16/IL-12+RT group than in B16 + RT group ( P &lt; 0.001)., Conclusions: During immune microenvironment reconstruction, overexpression IL-12 in the tumor microenvironment can reduce the percentage of PD-1 + T cells and suppress the growth of malignant melanoma in mice.

Objective: To investigate the effects of cytomegalovirus (CMV) DNA load on immune reconstitution and clinical outcomes of patients after unrelated cord blood transplantation (UCBT)., Methods: Eight-color flow cytometry was used to dynamically monitor the changes of peripheral blood lymphocyte subsets of 41 patients at one year after UCBT, and 10 healthy volunteers were enroled as controls. Patients were divided into two groups according to the DNA load of CMV (DNA copies <1000/ml and DNA copies ≥1000/ml). Comparative analyse of the effect of CMV DNA load on lymphocyte subsets and transplantation outcomes were carried out after transplantation., Results: The high CMV DNA load group showed a faster and expanded T cell reconstitution, and the differences between the two groups were statistically significant at one and nine months after transplantation (0.38×10 9 /L vs 0.25×10 9 /L, P=0.015 and 2.53×10 9 /L vs 1.36×10 9 /L, P=0.006, respectively). Further analysis of T cell subsets suggested that CD8 + T cells presented a higher and faster recovery in the high DNA load group, and the differences between the two groups were statistically significant at one and nine months after transplantation (0.20×10 9 /L vs 0.10×10 9 /L, P=0.038 and 1.62×10 9 /L vs 0.68×10 9 /L, P=0.003, respectively). In addition, there were no significant differences in levels of B cells, regulatory B cells and NK cells between the two groups. Outcomes after one- and a-half-year transplantation showed that there were no significant difference in relapse, non-relapse mortality and overall survival between the high and the low DNA load groups (7.7% vs 7.5%) (P=0.900) (15.4% vs 21.4%) (P=0.686) and (76.9% vs 78.6%) (P=0.889) respectively., Conclusion: The high CMV DNA load induces a faster and long-lasting expansion of T cells, mainly as the expansion of CD8 + T cells after UCBT. Besides, under the current pre-emptive treatment of CMV, the high CMV DNA load does not affect the early survival of patients with acute myeloid leukemia after UCBT.

Severe combined immunodeficiency disease (SCID) is a group of rare congenital diseases characterized by severe deficiencies in T lymphocyte counts and/or function. The recurrent, persistent and severe infections are its clinical manifestations. Neonatal screening and immune system reconstruction would improve the prognosis of SCID children. Newborn screening programs based on T-cell receptor excision circles (TRECs) quantitative detection have been carried out in clinical practice, however, the methods still have some limitations. Other new methods such as mass spectrometry and T lymphocyte-specific biomarker assays are still under investigation. Hematopoietic stem cell transplantation and gene therapy are the two main methods for reconstructing immune function in SCID children. Through improving the success rate of transplantation and the long-term safety and stability of viral vectors, some achievements have been made by many centers already. However, large-scale prospective studies are needed for evaluation of the long-term efficacy. In this article, the recent progress in newborn screening and immune reconstitution of SCID is reviewed.

Objective: To investigate the immune reconstruct regularity profile of KIR2DL1 and KIR3DL1 in unrelated-donor allogeneic hematopoietic stem cell transplantation (allo-HSCT) with KIR-AA genotype. Method: 75 donor-recipient pairs were performed by KIR genotying using PCR-SSP, and all donors were identified with KIR-AA genotype. Dynamic detections (including unrelated-donor on the day of transplantation and the recipient each month post allo-HSCT) of the expression of KIR2DL1/3DL1 on NK cell and mRNA level were performed in 291 cases using flow cytometry (FCM) and real-time fluorescent quantitation PCR (RT-qPCR) . Result: ①The median expression of KIR2DL1 in unrelated-donor on transplant's day was 21.60%, the median expression of KIR2DL1 in recipient 1M, 2M, 3M and 3-6M after transplantation were 7.40%, 12.00%, 16.92%, 17.64% respectively. The median expression of KIR2DL1 in unrelated-donor on transplant's day was 265.14 copies/10 000abl copies, the median expression of KIR2DL1 in recipient 1M, 2M, 3M, 3-6M, 6-9M, 9-12M after transplantation were 332.17, 438.31, 723.25, 414.17, 180.76 and 234.67 copies/10 000abl copies respectively. The median expression of KIR2DL1 on NK cells and mRNA level gradually increased at all time points after transplantation, and reached the highest expression at 3 months after transplantation. But mRNA expression levels increased earlier than NK cell membrane proteins. ②The median expression of KIR3DL1 in unrelated-donors on transplant's day was 18.56%, the median expression of KIR3DL1 in recipient 1M, 2M, 3M, 3-6M after transplantation were 23.83%, 22.57%, 23.02%, 21.60% respectively. The median expression of KIR3DL1 in unrelated-donor on transplant's day was 572.29 copies/10 000abl copies, the median expression of KIR3DL1 in recipient 1M, 2M, 3M, 3-6M, 6-9M, 9-12M after transplantation were 1 233.74, 1 140.42, 876.73, 1 057.07, 739.02 and 514.43 copies/10 000abl copies respectively. The median expression of KIR3DL1 on NK cells and mRNA level were higher than donors at 1 month after transplantation, and stable expression at all time points after transplantation, so mRNA and NK cell membrane proteins expression increased at the same time. Conclusion: The immune reconstruct regularity of KIR2DL1 and KIR3DL1 gene were different, which provided an experimental basis for selecting the best time to detect the expressions of KIR2DL1 and 3DL1 after transplantation.