| Background Delayed immune reconstitution (IR) is usual in allogeneic hematopoietic stem cell transplantation (allo-HSCT), especially for CD4+helper T cells, which is one important cause of infections, relapse and secondary tumors. Invasive fungal infections (IFI) can cause significant mortality after allo-HSCT. Recent studies showed that four subsets of Th cells including Th17(T helper17), Th1(T helper1), Th2(T helper2) and regulatory T cells (Treg) play different roles in immune responses to IFI. Thl and Th17cells can help to resist IFI while Th2and Treg cells can suppress immune responses to IFI. Immune reconstitution characteristics of these Th subsets after allo-HSCT have been rarely studied and there has been no report about their associations with IFI in allo-HSCT.Objective We aim to study the IR characteristics of CD4+Th cells and its four subsets during first three months after allo-HSCT and investigate their relationships with IFI and further study their roles in occurrence of IFI.Methods Peripheral blood samples of152detections from62patients undergoing allo-HSCT from July2013to March2014were collected at14days,1month,2months,3months post transplant. Healthy controls (HC) were26healthy donors. Flow cytometry was used to analyze ratio of T17, Thl, Th2and Treg in CD4+T cells. Real time PCR (RT-PCR) was used to detect mRNA expression level of their specific transcription factors such as retinoic acid-related orphan receptor y (RORyt), T box expressed in T cells (T-bet), GATA3(GATA binding protein-3), forkhead/winged helix transcription factor P3(FoxP3). ELISA was used to measure plama concentrations of interferon-y (IFN-y), interleukin6(IL-6), interleukin10(IL-10), transforming growth factor-β (TGF-β). Factors affecting IR were analyzed and groups were then divided into subgroups according to these factors. These factors were compared among patients detected at1month,2months,3months post transplant and HC respectively. The changes of these factors at different months for the same patients were studied. Logistic regression analysis was used to study the relationships between CD4+T cell related factors and IFI.Results:1Factors affecting IR after allo-HSCT CD3+cells count at1month post transplant was associated with recipient age (β=-0.308, P=0.039), CD4+T cells count at1month post transplant was associated with aGVHD history (β=-0.355, P=0.017), donor source ((β=-0.427, P=0.046), recipient age (β=-0.365, P=0.009). CD3+cells count at2months post transplant was associated with antithymocyte globulin (ATG) conditioning (β=-0.387, P=0.021). CD4+T cells count at3months post transplant was associated with ATG conditioning (p=-0.387, P=0.043).2Comparisions of each factor among patients detected at different months post transplant and HC (1) CD3+cells count at1month post transplant [(559.93±323.27)×106/L, P=0.000] was lower than HC [(1311.55±363.68)×106/L]. CD3+cells count at2months post transplant had no significant difference between non-ATG conditioning group [(1463.62±900.71)×106/L] and HC [(1311.55±363.68)×106/L](P=0.661), but it was lower in ATG conditioning group [(798.33±602.58)×106/L] than HC (P=0.001) and non-ATG conditioning group (P=0.002). CD3+cells count had no significant difference between patients detected at3months post transplant and HC. CD4+T cells count at1month post transplant was lower for all subgroups including SD-nonGVHD, SD-GVHD, URD-nonGVHD and URD-GVHD than HC. CD4+T cells count at2months post transplant [(458.40±364.47)×106/L] was lower than HC [(971.91±336.10)×106/L](P=0.000). CD4+T cells count of patients with ATG conditioning [(300.73±224.75)×106/L, P=0.0001and non-ATG conditioning [(537.70±221.88)×106/L, P=0.0001were both lower than HC [(971.91±336.10)×106/L] at3months post transplant while there was no significant difference between both of them (P=0.057). Compared with HC [(339.64±122.52)×106/L], CD8+T cells count was lower at1month post transplant [(252.13±184.72)×106/L, P=0.004] while higher in both2months [(763.30±576.59)×106/L, P=0.003] and3months [(633.45±97.71)×106/L, P=0.002] post transplant. Th17、Thl、Th2、Treg count were all lower than those of HC (P<0.05).(2) The ratio of Thl7cells in CD4+cells was all lower than HC at1month,2months and3months post transplant (P<0.05). The ratio of Thl cells had no significant difference between HC and1month or2months post transplant while it was higher at3months post transplant [(28.54±14.69)%] than HC [(20.74±10.26)%](P=0.034). The ratio of Th2cells was lower at1month post transplant [(0.22±0.20)%] than HC [(0.50±0.57)%](P=0.009) while there was no significant difference between HC and2or3months post transplant. The ratio of Treg cells had no significant difference between HC and1month,2or3 months post transplant.(3) The mRNA levels of RORyt、GATA3、FoxP3were all lower at1month,2months and3months post transplant than HC (P<0.05). The mRNA expression of T-bet was lower at both1month (0.3111±0.3513, P=0.000) and3months (0.4666±0.3960, P=0.045) post transplant than HC (0.7279±0.5477) while there was no significant difference between2months post transplant and HC.(4) Plasma concentration of IFN-y had no significant difference between HC and1month,2or3months post transplant. The concentration of IL-6、IL-10was higher at1month,2months and3months post transplant than HC respectively (P<0.05). The concentration of TGF-β was all lower at1month,2months and3months post transplant than HC(P <0.05).3Changes of CD4+T cells subsets related factors among different months (1) Th17cells count was lower at1month [(2.49±2.16)×106/L] than3months post transplant [(3.95±2.91)×106/L, P=0.017] and there was no significant difference between1month and2months,2months and3months post transplant. Thl cells count was lower at1month [(78.88±88.89)×106/L] than3months post transplant [(139.52±84.08)×106/L](P=0.042] and there was no significant difference betweenl month and2months,2months and3months post transplant. There was no significant difference for Treg and Th2cells count comparisions among1month,2months and3months post transplant one another.(2) There was no significant difference for ratios of Thl7, Thl, Th2, Treg in CD4+T cells among1month,2months and3months post transplant respectively.(3) The mRNA expression of RORyt, T-bet, GATA3, FoxP3had no significant difference among1month,2months,3months post transplant.(4) There was no significant difference for concentration of IFN-y, IL-10, TGF-β among1month,2months and3months post transplant. The concentration of IL-6was lower at1month [(4.17±4.80)pg/ml] than3months post transplant [(13.87±10.36)pg/ml](P=0.005) while there was no significant difference between1month and2months,2months and3months comparisons.4Associations between CD4+T cells subsets and IFI (1) Among129detections of59cases, IFI occurred in12patients (20.3%), including1probable IFI case with aspergillus infection, and11possible IFI cases.(2) Thl7cells count was associated with occurrence of IFI (OR=0.176, P=0.005) while Thl, Th2and Treg cells count was not associated with IFI.(3) None of RORyt, T-bet, GATA3and FoxP3mRNA expression levels was associated with IFI.(4) None of plama concentrations of IFN-y, IL-6, IL-10, TGF-β was associated with IFI.Conclusions1Reconstitution of CD4+T cells and its subsets Th17, Thl, Th2, Treg is all delayed while CD8+T cells show rapid reconstitution in allo-HSCT.2Polarization of CD4+T cells to Th17cells is obviously inhibited after allo-HSCT.3Th17is a protective factor for occurrence of IFI after allo-HSCT. Objective To investigate the risk factors of intestinal acute graft-versus-host disease (aGVHD) after allogeneic hematopoietic stem cell transplantation (allo-HSCT).Methods The clinical data of534cases of533patients undergoing allo-HSCT during Jan2004and Sep2012were retrospectively analyzed. The effects of donor-recipient HLA mismatching, recipient age, donor age, donor-recipient sex combinations, donor-recipient relationship, stem cell source, conditioning regimen with or without total body irradiation (TBI) and HLA loci on intestinal aGVHD with different severity were analyzed by Logistic regression.Results Intestinal aGVHD occurred in123cases (23.0%), with86cases (16.1%) of stage1intestinal aGVHD (16.1%) and37cases (6.9%) of stage2to4intestinal aGVHD. Multivariate analysis showed that donor-recipient HLA mismatching (OR=2.519, P=0.002), increasing donor age (OR=1.034, P=0.003), female donor for male recipient (OR=1.855, P=0.007) were risk factors for intestinal aGVHD, HLA-B38(OR=0.256, P=0.032) was its protective factor. Donor-recipient HLA mismatching (OR=2.799, P=0.011), increasing donor age (OR=1.045, P=0.012), HLA-A1(OR=A.157, P=0.002), A30(OR=3.143, P=0.005) were risk factors for stage2to4intestinal aGVHD.Conclusions Occurrence of intestinal aGVHD and its severity are associated with donor-recipient HLA mismatching, donor age, donor-recipient sex relationships and some HLA loci. |
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