Interrelationship between immunocompetent and structural cells in post-burn scars

It has been suggested that immunological factors play a major role in the pathogenesis of hypertrophic scars. Characteristically there is an onset of typical clinical features and, after a variable period of activity, there is a phase of remission. The factors which produce this progressive change in the scars are not clear, and it is not known if their manipulation could provide a therapeutic progress. In order to further investigate the pathophysiology, morphological studies have been performed. In active hypertrophic scars lymphocytic infiltrates are abundant. Among them activated T-cells represent 70% of infiltrates while in normotrophic scar activated T-cells make up 30% of lymphocytes, suggesting a role of these cells in the mechanisms leading to scar hypertrophy. Upon activation, the lesional T-cells release several cytokines which may induce anomalous expression of activation markers (HLA-DR, ICAM-1, CD36, IL-2R) on keratinocytes and fibroblasts of hypertrophic tissue. A wide range of cytokines has been examined: among those analyzed the only one that changes in the remission phase is IFN. In fact, IFN is highly expressed in lymphocytes in active hypertrophic scars while it is less expressed in the remission phase and in control samples.Presented at the European Burns Association State of the Art Symposium, Control of the Burn Scar, Amsterdam, The Netherlands, October 4–5, 1996     

Effect of in vitro mechanical compression on Epilysin (matrix metalloproteinase-28) expression in hypertrophic scars

Epilysin, designated matrix metalloproteinase (MMP)-28, is the newest member of this family of proteases expressed by keratinocytes in response to an injury. MMP-28's physiological role and specific substrates are unknown, but its expression pattern suggests that it may serve a role in both tissue homeostasis and wound healing. The aim of this preliminary study was to observe the presence of MMP-28 protein in normotrophic and hypertrophic scars and to evaluate the effect of in vitro mechanical compression on its expression. Biopsies from normotrophic and hypertrophic scars resulting from burns were divided into two samples, one to be used as control (uncompressed) and the other to be compressed in an oxygenated organ chamber for 24 hours in the presence of a serum-free medium, using an electromechanical load transducer (stable pressure = 35 mmHg). Analysis of MMP-28 protein secretion, assessed by Western blot and beta-casein zymography in scar conditioned media, revealed that normotrophic scar did not release MMP-28 in any condition while hypertrophic scar released active MMP-28 both in control conditions and after compression. MMP-28 immunohistochemistry revealed a light protein presence in normotrophic scar keratinocytes and a strong MMP-28 positivity in hypertrophic scar keratinocytes in control conditions, while compression increased MMP-28 staining in normotrophic scar and induced a significant reduction of the protein presence in hypertrophic scar keratinocytes. As it has been suggested that MMP-28 may restructure the skin basal membrane (Saarialho-Kere et al., 2002), our data indicate that mechanical compression directly acts to modulate the remodeling phase of wound healing, altering release and activity of MMP-28 in hypertrophic scars.     

Functional analysis of T lymphocytes infiltrating the dermis and epidermis of post-burn hypertrophic scar tissues

The cytokine profile of T cell clones (TCC) from the dermis and epidermis of burn patients with hypertrophic scars (HS) in active (AHS) and remission phases (RHS) was determined in this study. We found that AHS tissues are heavily infiltrated by Type 0-Type 1 polarized CD3+ lymphocytes producing high IFN-gamma and low IL-4 levels. Analysis of their surface marker phenotype showed that the high IFN-gamma production was shared equally between the CD4+ TCRalpha/beta and CD8+ TCRalpha/beta clones. The profile of TCC from RHS tissues revealed pronounced infiltration of Type 0-Type 1 polarized lymphocytes with an even more evident Type 1 profile. However, the levels of IFN-gamma produced by RHS-derived TCC were 4-6 times lower than those produced by AHS-derived TCC. These data show that high levels of IFN-gamma produced by Type 0-Type 1 lymphocytes infiltrating HS are a feature of AHS, whereas reduction of this ability to produce high levels of IFN-gamma, though without a shift towards a Type 0-Type 2 phenotype through an increase in IL-4, is characteristic of RHS.     

Nerve outgrowth and neuropeptide expression during the remodeling of human burn wound scars. A 7-month follow-up study of 22 patients

Increasing data suggest that the skin nerve system is involved in wound healing. The objective of this study was to investigate the outgrowth of nerve fibers during the burn wound remodeling process and to analyze possible differences between normotrophic and hypertrophic burn wounds. In a prospective study, biopsies were taken from 22 patients with spontaneously healed partial-thickness burns at 1, 4 and 7-month post-burn. Nerve outgrowth and the expression of the neuropeptides substance P, neurokinin A, calcitonin gene-related peptide, vasoactive intestinal peptide and neuropeptide Y was monitored using immunohistochemistry. Our results showed that the number of nerve fibers gradually increased in both the dermis and the epidermis, but that they did not reach the levels of expression present in matched unburned skin of the same patient. A significantly higher number of nerve fibers were observed in normotrophic scars compared with hypertrophic scars. The number of neuropeptides-containing nerves in normotrophic and hypertrophic scars were similar. In conclusion: 7 months after wound closure, burn wound scars contain less nerve fibers than unburned skin. The significantly higher number of nerve fibers in normotrophic, compared with hypertrophic scars suggests a regulatory role for the skin nerve system in the outcome of burn wound healing.     

48例成人慢性活动性EB病毒感染的临床特征和外周血淋巴细胞免疫表型

目的探讨成人慢性活动性EB病毒感染（CAEBV）患者的临床特征及外周血淋巴细胞免疫表型。 方法回顾性纳入2017年3月至2019年3月如皋市人民医院收治的48例成人CAEBV感染者,记录患者一般临床资料及EB病毒相关抗体、病毒载量和细胞免疫表型。 结果48例成人CAEBV患者年龄（40.8 ± 14.4）岁,均有发热症状。严重肝功能衰竭、Hermansky-Pudlak综合征（HPS）、中枢神经系统（CNS）病变和间质性肺炎发生率分别为29.2%（14/48）、47.9%（23/48）、12.5%（6/48）和22.9%（11/48）。患者血浆EBV DNA拷贝数中位数为2.9 × 10⁴（5 × 10²～5.8 × 10⁶）拷贝/ml。32例患者完成外周血淋巴细胞免疫表型检测,B细胞、NK细胞、CD4⁺ T细胞、CD8⁺ T细胞水平分别为25 × 10⁹/L（0,19.6 × 10⁹/L）、82.5 × 10⁹/L（54 × 10⁹/L,183 × 10⁹/L）,332.5 × 10⁹/L（178.25 × 10⁹/L,534.25 × 10⁹/L）,302 × 10⁹/L（190 × 10⁹/L,573.5 × 10⁹/L）,53.1%（25/48）患者出现CD4/CD8比值倒置（< 1）。CD38⁺CD8⁺ T细胞百分比中位数为93.2%（49.1%,99.7%）,HLA-DR⁺CD8⁺T细胞百分比的中位数为71.25%（17.1%,97.5%）。 结论成人CAEBV患者的临床特征与其外周血淋巴细胞免疫表型关系密切,B细胞、NK细胞和CD4⁺ T细胞水平多降低。     

Differences in collagen architecture between keloid, hypertrophic scar, normotrophic scar, and normal skin: An objective histopathological analysis

Normotrophic, hypertrophic, and keloidal scars are different types of scar formation, which all need a different approach in treatment. Therefore, it is important to differentiate between these types of scar, not only clinically but also histopathologically. Differences were explored for collagen orientation and bundle thickness in 25 normal skin, 57 normotrophic scar, 56 hypertrophic scar, and 56 keloid biopsies, which were selected on clinical diagnosis. Image analysis was performed by fast fourier transformation. The calculated collagen orientation index ranged from 0 (random orientation) to 1 (parallel orientation). The bundle distance was calculated by the average distance between the centers of the collagen bundles. The results showed that compared with all three types of scars, the collagen orientation index was significantly lower in normal skin, which indicates that scars are organized in a more parallel manner. No differences were found between the different scars. Secondly, compared with normal skin, normotrophic scar, and hypertrophic scar, the bundle distance was significantly larger in keloidal scar, which suggests that thicker collagen bundles are present in keloidal scar. This first extensive histological study showed objective differences between normal skin, normotrophic, hypertrophic, and keloidal scar.     

Tenascin-C在瘢痕疙瘩和增生性瘢痕中的基因表达研究

目的 探讨Tenascin-C基因在瘢痕疙瘩和增生性瘢痕中的表达。方法 取正常成人皮肤组织RNA，构建正义、反义Tenascin-C(Tn-C)mRNA探针，运用原位杂交技术，观测10例瘢痕疙瘩、10例增生性瘢痕和5例正常成人皮肤组织中Tn-C mRNA的表达。结果 Tn-C mRNA在正常皮肤表皮中无表达，真皮中表达稀少，局限于乳头真皮层的成纤维细胞和皮肤附属器；10例瘢痕疙瘩表皮均有表达，真皮分布较广，如成纤维细胞、血管内皮和皮肤附属器；Tn-C mRNA在3例增生性瘢痕表皮表达，7例无表达，真皮中表达与瘢痕疙瘩相同但较弱，比正常皮肤增多，但差异无显著性。结论 Tenascin-C mRNA在瘢痕疙瘩表皮和真皮中有高表达。     

Some observations on the surface structure of collagen in hypertrophic scars

The structure of the interface between epidermis and dermis was examined in normal skin and in hypertrophic scars using the scanning electron microscope. The hypertrophic scar was found to show a completely different structure of fibrous tissue (collagen) at this surface when compared to normal skin. As the hypertrophic scar matured this surface was remodelled to resemble more closely the surface observed in normal skin. The study suggests that the attachment of epidermal cells to the surface of the fibrous tissue developing in the burn wound may be an important aspect of the pathogenesis of hypertrophic scarring.     

Proteoglycan synthesis in human skin and burn scar explant cultures

The synthesis of proteoglycans (PG) by normal human skin, and normal and hypertrophic scars were compared using tissue explants in culture. Newly synthesized PG were labelled with [³⁵S]Na₂SO⁴. Significant differences were found in the proportion of [³⁵S]-radio-labelled incorporation of PG in the tissue and accumulation of [³⁵S]PG in culture medium in the different tissues. The rate of PG biosynthesis in all three tissue types occurred in two phases. There was an initial phase of PG synthesis occurring at 0–3 h and a later phase that occurred at 3–18 h [³⁵S]-labelled PG were isolated and characterized by Sepharose CL-6B chromatography and cellulose acetate electrophoresis. The results showed that the hypertrophic scar tissue and its culture medium contained higher proportions of dermatan sulphate (DS), chondroitin sulphate (CS) and DS' PG than the normal skin fractions. These results suggest that abnormal scarring is related to a change in the level of PG synthesis during the burn injury repair process.     

Immunohistochemical analysis of neuropeptides (protein gene product 9.5, substance P and calcitonin gene-related peptide) in hypertrophic burn scar with pain and itching

BackgroundNeuropeptides have been recently reported as having an important role in wound repair, and relief from pain and itching sensation. The aim of this study was to evaluate the effect of neuropeptides on the wound healing process in hypertrophic scar formation that accompanies severe pain and itching sensation.MethodsWe collected forty-three hypertrophic scar specimens from hypertrophic scar release and skin graft under general anesthesia. Immunohistochemical stains for protein gene product (PGP) 9.5, substance P (SP), and calcitonin gene-related peptide (CGRP) were performed. Pain and itching over the scar were recorded using verbal numerical rating scale (VNRS).ResultsIn the epidermis, PGP 9.5, SP, and CGRP were significantly increased in hypertrophic scars compared with matched unburned skin. In the reticular dermis, SP and CGRP were significantly increased in hypertrophic scars compared with control. The pain and itching verbal numerical rating scale in scar group were significantly higher compared to control. In the papillary dermis, the PGP represented significant correlation with Itching P (correlation coefficient 0.698) and the SP represented significant correlation with pain N (correlation coefficient −0.671). In the reticular dermis, the SP represented significant correlation with pain N (correlation coefficient −0.614) and CGRP represented significant correlation with pain P/Itching P (correlation coefficient 0.801/0.611).ConclusionsNeuropeptides such as PGP 9.5, SP, and CGRP seem to affect scarring via sensory neurotransmission, it have a regulatory role for pain and itching sensation in hypertrophic scars.     

手足口病患儿外周血CXC趋化因子配体10表达、T细胞及细胞因子与预后的相关性

目的检测手足口病（HFMD）患儿外周血中的CXC趋化因子配体10（CXCL-10）表达水平、T淋巴细胞亚群及相关细胞因子与HFMD患儿预后的相关性。 方法选取2016年4月至2017年5月徐州市儿童医院收治的HFMD患儿60例为研究对象,根据病情诊断标准分为重症组（10例）和轻症组（50例）;根据随访结果又分为预后良好组（58例）和预后不良组（2例）。选择同期体检正常婴幼儿52例为对照组。采用ELISA法检测两组研究对象外周血CXCL-10表达水平;采用流式细胞仪检测外周血T淋巴细胞亚群,采用ELISA法检测外周血中白细胞介素-17（IL-17）、IL-22、IL-23以及肿瘤坏死因子-α（TNF-α）水平。 结果重症组患儿外周血CXCL10水平、IL-17、IL-22、IL-23和TNF-α水平显著高于对照组和轻症组患儿（P均< 0.001）。轻症组患者细胞因子CXCL-10、IL-17、IL-22、IL-23和TNF-α水平高于对照组（P均< 0.005）。重症组HFMD患儿外周血CD3⁺ T、CD4⁺ T、CD8⁺ T以及CD4⁺/CD8⁺淋巴细胞比例显著低于轻症组患儿和对照组（P均< 0.05）;轻症组患儿外周血CD3⁺ T、CD4⁺ T、CD8⁺ T、CD4⁺/CD8⁺淋巴细胞比例与对照组差异有统计学意义（P均< 0.05）;经Pearson相关系数分析,HFMD患儿外周血CXCL-10表达水平与T淋巴细胞亚群CD3⁺T、CD4⁺ T、CD8⁺ T、CD4⁺/CD8⁺表达水平均负相关（r =－0.609、－0.714、－0.514、－0.524,P = 0.014、0.023、0.001、0.006）,HFMD患儿外周血中IL-17、IL-22、IL-23和TNF-α水平与外周血CXCL-10水平正相关（r = 0.519、0.473、0.418、0.459,P = 0.002、0.006、0.009、0.007）;预后不良组患儿外周血CXCL-10表达水平显著高于预后良好组（t = 2.055、P = 0.044）,外周血CD3⁺ T、CD4⁺ T、CD8⁺ T淋巴细胞比例显著低于预后良好组,差异均有统计学意义（t = 2.508、P = 0.015,t = 3.830、P < 0.001,t = 2.222、P = 0.030）。 结论手足口病患者外周血CXCL-10、T淋巴细胞亚群检测对病情监测及判断预后有重要价值。     

肺移植术后T细胞亚群研究进展

T细胞是肺移植术后免疫应答的主要效应细胞,其细胞亚群水平对肺移植受者机体免疫状态具有重要影响。本文综述CD4^＋ T细胞、CD8^＋ T细胞和调节性T细胞的免疫学机制及其与肺移植术后原发性移植肺功能障碍、排斥反应、免疫耐受和感染等的关系,同时探讨监测肺移植术后T细胞亚群的临床意义。     

病理性瘢痕中Langerhans细胞的形态学研究

目的：探讨Langerhans细胞与病理性瘢痕的相关性.方法：应用免疫组织化学法（ABC法）观察瘢痕疙瘩、增生性瘢痕及正常皮肤组织中S-100蛋白和CD1a的表达,用透射电镜观察这三种组织中的Langerhans细胞的超微结构.结果：瘢痕疙瘩及增生性瘢痕组织中对S-100蛋白和CD1a免疫反应阳性的Langerhans细胞较正常皮肤中明显增多.瘢痕疙瘩及增生性瘢痕与正常皮肤之间有显著性差异（P＜0.05）.瘢痕疙瘩与增生性瘢痕之间差异不显著（P＞0.05）.电镜下表皮细胞层中Langerhans细胞核形多不规则,胞质中等电子密度,有较多线粒体和溶酶体,扁平囊泡较多.结论：Langerhans细胞在瘢痕疙瘩及增生性瘢痕组织中明显增多,且功能活跃.     

甲型流感病毒感染者CD160 +CD8 +T淋巴细胞亚群分析

目的研究甲型流感病毒（IAV）感染者外周血中CD160⁺CD8⁺ T淋巴细胞亚群的特征。 方法收集2018年12月至2019年3月流感季就诊于北京大学地坛医院教学医院感染二科IAV感染者共37例及同期体检的健康对照者51例,应用多色流式细胞检测平台,检测外周血不同分化阶段CD160⁺CD8⁺ T淋巴细胞的比例。 结果与健康对照相比,IAV感染者外周血T细胞中CD160⁺CD8⁺ T细胞比例下降[24.50%（14.70%,44.10%）vs. 8.85%（5.14%,18.15%）],差异有统计学意义（U = 326.00、P < 0.001）。IAV感染者与健康对照相比,其CD8⁺ T中初始T细胞（T_N）[3.88%（1.28%,9.48%） vs. 0.22%（0.12%,0.60%）]、中央记忆性T细胞（T_CM）[8.70%（4.43%,15.80%） vs. 1.51%（0.69%,2.54%）]、效应记忆性T细胞（T_EM）[22.30%（13.80%,30.40%） vs. 7.71%（5.23%,16.25%）]和终末效应记忆T细胞（T_EMRA）[（46.99 ± 22.91）% vs. （21.60 ± 13.38）%]4个亚群中CD160⁺细胞的比例均下降,差异有统计学意义（U = 238.50、81.50、412.00,t = 6.03;P均< 0.001）。IAV感染者出院时较入院时CD160⁺CD8⁺ T细胞比例上升[（8.92 ± 7.84）% vs. （16.40 ± 9.43）%]（t = 4.09、P = 0.001）,但仍低于正常水平[24.50%（14.70%,44.10%） vs. （16.40 ± 9.43）%],差异有统计学意义（U = 209.50、P = 0.0029）。 结论甲型流感病毒感染者外周血中T淋巴细胞CD160⁺CD8⁺ T细胞亚群比例显著降低,可能是免疫失衡的重要表现。     

肾移植受者T细胞亚群绝对计数动态监测对感染的预警作用

目的探讨长期监测T细胞亚群绝对计数水平对肾移植受者术后感染的预警作用。 方法回顾性分析2017年1月至2021年5月在上海交通大学医学院附属瑞金医院26例行肾移植术后新发感染受者临床资料(感染组,感染发生在移植后1～240个月)。选择129例同期肾移植术后无感染、健康受者作为对照组。感染组连续或定期测量外周血T细胞亚群CD3^＋、CD4^＋和CD8^＋绝对计数,并与对照组检测数据进行比较。根据移植后采样时间将感染组和对照组各分为6个亚组,分析感染亚组与其相应对照亚组之间T细胞亚群绝对计数的差异。正态分布计量资料采用两独立样本t检验和单因素方差分析比较,非正态分布计量资料采用Mann-Whitney U检验比较,计数资料采用χ²检验比较。使用受试者工作特征(ROC)曲线分析T细胞亚群绝对计数在肾移植术后预警感染性疾病的最优值。P<0.05为差异有统计学意义。 结果感染组和对照组受者CD4^＋/CD8^＋比值分别为(1.2±0.5)、(1.3±0.6),差异无统计学意义(t＝0.610,P>0.05)。感染组受者CD3^＋、CD4^＋和CD8^＋ T细胞绝对计数[(367±212)、(189±117)和(161±92)个/μL]均低于对照组[(1 374±663)、(695±334)和(626±377)个/μL],差异均有统计学意义(t＝14.036、13.541和12.311,P均<0.05)。CD3^＋、CD4^＋和CD8^＋ T细胞绝对计数在6个感染亚组受者中差异均无统计学意义(P均>0.05)。对照亚组1受者CD3^＋、CD4^＋和CD8^＋ T细胞绝对计数均低于对照亚组5,差异均有统计学意义(P均<0.05)。CD4^＋、CD8^＋和CD3^＋绝对计数预测肾移植术后感染性疾病最优截断值分别为712、362和255个/μL,敏感度分别为94.6%、92.2%和96.1%,特异度分别为92.3%、96.2%和88.5%。 结论肾移植受者低T细胞亚群绝对计数水平具有预示及预警感染风险的作用。     

感染性肺炎新生儿免疫功能系统评价及临床意义

目的探讨感染性新生儿肺炎免疫功能变化及其临床意义。 方法选取2016年3月至2017年10月深圳市龙华区中心医院新生儿科收治的感染性新生儿肺炎60例为观察组,同期选取健康新生儿60例为对照组。根据肺炎严重程度将观察组分为两个亚组：轻症组（39例）和重症组（21例）;根据新生儿感染性肺炎分期差异分为两个亚组：急性期组（24例）和恢复期组（36例）。入组患儿均于急性期、恢复期采集血样,用于体液免疫指标（免疫球蛋白IgA、IgM、IgG1、IgG2、IgG3、IgG4及补体C3、C4水平）和细胞免疫指标（CD3⁺ T、CD4⁺ T、CD8⁺ T、CD4/CD8、NK细胞）检测。 结果重症组患儿血清IgA、IgG1、IgG2、IgG3、IgG4水平、CD3⁺ T细胞、CD4⁺ T细胞、NK细胞比例及CD4/CD8较对照组和轻症组患儿均显著降低（P均< 0.05）,IgM、C3、C4水平、CD8⁺ T细胞比例较对照组和轻症组患儿均显著升高（P均< 0.05）,差异均有统计学意义。Spearman相关分析发现,血清IgA、IgG1、IgG2、IgG3、IgG4水平、CD3⁺ T细胞、CD4⁺ T细胞、NK细胞比例及CD4/CD8与新生儿感染性肺炎严重程度呈负相关（r =－0.826、－0.826、－0.665、－0.822、－0.826、－0.816、－0.794、－0.824、－0.820,P均< 0.001）;血清C3水平、CD8⁺ T细胞比例与新生儿感染性肺炎严重程度呈正相关（r = 0.467、0.788,P均< 0.001）。急性期患儿血清IgA、IgG1、IgG2、IgG3、IgG4水平、CD3⁺ T细胞、CD4⁺ T细胞、NK细胞比例及CD4/CD8均较恢复期和对照组显著降低（P均< 0.05）,而血清IgM、C3、C4水平、CD8⁺ T细胞均较恢复期和对照组显著升高,差异均有统计学意义（P均< 0.05）。 结论感染性新生儿肺炎细胞免疫和体液免疫功能下降,与疾病严重程度呈负相关,且疾病不同阶段对免疫功能影响较大,应对该类患儿免疫功能加强调控。     

系统性红斑狼疮患者不同巨细胞病毒感染状态淋巴细胞亚群分析

目的探讨系统性红斑狼疮（SLE）患者不同巨细胞病毒（CMV）感染状态淋巴细胞亚群的特征，为临床诊断系统性红斑狼疮提供依据。 方法选取2016年6月至2019年6月于南京中医药大学附属张家港医院的SLE住院患者共96例，其中合并巨细胞病毒血症患者18例（巨细胞病毒血症组）、巨细胞病毒病患者50例（巨细胞病毒病组）和无CMV感染者28例（对照组）。获取3组患者的一般资料、常规实验室指标、CMV DNA拷贝数和外周血淋巴细胞亚群计数；并比较CMV感染者的上述指标，分析不同CMV感染状态下淋巴细胞亚群的特征。 结果巨细胞病毒血症组患者和巨细胞病毒病组患者的红细胞沉降率（t =-0.141、P = 0.025，t =-0.194、P = 0.003）CRP水平显著高于对照组（t =-0.563、P = 0.010），巨细胞病毒病组患者CRP（t =-0.854、P = 0.006）、环磷酰胺治疗例数（χ² =-6.139、P = 0.013）、血CMV DNA拷贝数（t =-0.355、P = 0.041）均显著高于巨细胞病毒血症组。巨细胞病毒血症组患者与对照组患者总淋巴细胞计数、CD3⁺ T细胞、CD3⁺CD4⁺ T细胞、CD3⁺CD8⁺ T细胞、CD19⁺ B细胞和CD56⁺CD16⁺ NK细胞计数差异均无统计学意义（P均> 0.05）。巨细胞病毒病患者总淋巴细胞计数（t = 0.933、P < 0.001）、CD3⁺ T细胞（t = 0.177、P = 0.018）、CD3⁺CD4⁺ T细胞（t = 0.207、P < 0.001）、CD3⁺CD8⁺ T细胞（t = 0.169、P < 0.001）和CD19⁺ B细胞（t = 0.320、P = 0.023）显著低于对照组患者。 结论淋巴细胞计数减低（尤其是CD4⁺ T细胞计数减低）常见于SLE患者伴发CMV感染中常见，是系统性红斑狼疮患者CMV感染诊断的潜在生物标记物。     

宫颈上皮内病变患者人乳头状瘤病毒感染与Th17细胞、Treg细胞及相关细胞因子表达的相关性

目的探讨宫颈上皮内病变（CIN）患者人乳头状瘤病毒（HPV）感染与Th17细胞、Treg细胞及相关细胞因子表达的相关性。 方法选择2017年2月至2018年9月于新疆自治区人民医院就诊的CIN患者共140例进行回顾性分析,根据是否感染HPV分为感染组（86例）和非感染组（54例）,采用免疫组织化学方法比较两组患者T细胞CD3、CD4及CD8水平以及Treg和Th17的表达水平,采用流式细胞技术比较两组患者的外周血T细胞亚群含量（CD3⁺ T细胞、CD4⁺ T细胞、CD8⁺ T细胞的百分比）、Treg细胞和Th17细胞百分含量（CD4⁺CD25⁺ Treg、CD8⁺CD25⁺CD127^-Treg、CD4⁺Foxp3⁺ Treg、CD8⁺CD25⁺Foxp3⁺ Treg和CD4⁺IL17⁺Th17以及Th17/Treg比值）以及相关细胞因子[肿瘤坏死因子（TGF）-β、白细胞介素（IL）-10、IL-17和干扰素（IFN）-γ]含量的差异,应用多因素Logistic回归模型分析CIN患者感染HPV的危险因素。 结果免疫组织化学结果显示,两组患者CIN组织中T细胞CD3、CD4和CD8的表达水平差异无统计学意义,感染组患者CIN组织中T细胞抗-FOX3（Treg）和Th17表达水平显著高于非感染组。流式细胞术检测结果显示两组患者CD3⁺ T细胞、CD4⁺ T细胞、CD8⁺ T细胞百分比差异无统计学意义（t = 0.870、1.509、0.236;P = 0.385、0.133、0.813）,感染组患者CD4⁺CD25⁺ Treg、CD8⁺CD25⁺CD127^—Treg、CD4⁺Foxp3⁺ Treg、CD8⁺CD25⁺Foxp3⁺ Treg、CD4⁺IL17⁺ Th17以及Th17/Treg比值均显著高于非感染组（P均< 0.05）。感染组患者外周血TGF-β、IL-10和L-17均高于非感染组（t =－11.601、－42.251、－40.31,P均< 0.001）,而IFN-γ低于非感染组（t = 10.316、P < 0.001）。多因素Logistic回归分析显示,CD4⁺CD25⁺ Treg（OR = 3.673、P = 0.031）、CD4⁺IL17⁺Th17（OR = 1.974、P = 0.021）和Th17/Treg（OR = 3.585、P = 0.024）均为CIN患者感染HPV的危险因素,IFN-γ（OR = 0.612、P = 0.028）为CIN患者感染HPV的保护因素。 结论CIN患者HPV感染与Treg细胞、Th17细胞及IFN-γ因子存在相关性,HPV感染的CIN患者Treg和Th17细胞增高,IFN-γ降低。     

Increased TGF-β–producing CD4+ T lymphocytes in postburn patients and their potential interaction with dermal fibroblasts in hypertrophic scarring

The development of hypertrophic scar involves a complex interplay between cells and cytokines. Although the mechanism underlying its pathogenesis is not well understood, a polarized T-helper type 2 immune response has been reported, indicating a role for CD4+ T lymphocytes in hypertrophic scarring. Here, we report an increased frequency of CD4+/transforming growth factor-beta (TGF-beta)-producing T cells in the peripheral blood and hypertrophic scar tissue of burn patients. These cells may play an indirect regulatory role in hypertrophic scar by affecting the functions of dermal fibroblasts. Our results show an increase in cell proliferation and collagen synthesis by dermal fibroblasts treated with medium derived from burn patient CD4+ T lymphocytes but not from the CD4+ T cells of normal subjects. Using confocal microscopy and immunoblotting, we found the level of alpha-smooth muscle actin to be elevated in these treated dermal fibroblasts, which also showed an enhanced ability to contract collagen lattices. TGF-beta levels in medium conditioned by the culture of CD4+ T lymphocytes from burn patients were significantly higher than in the conditioned medium from CD4+ T lymphocytes of normal subjects. In addition, the application of a TGF-beta-neutralizing antibody significantly reduced the effect of burn patient CD4+ T lymphocyte medium on dermal fibroblast proliferation and collagen lattice contraction. Our study suggests that CD4+/TGF-beta-producing T lymphocytes may play an important role in postburn hypertrophic scarring.