反复呼吸道感染儿童血清维生素A、维生素E水平与免疫球蛋白、T淋巴细胞亚群、NK细胞及骨密度的关系分析

摘要 目的：探讨反复呼吸道感染（RRTI）儿童血清维生素A、维生素E水平与免疫球蛋白（Ig）、T淋巴细胞亚群、NK细胞及骨密度的关系。方法：选择2018年2月至2020年12月我院儿科收治的107例RRTI患儿（感染组）和83例同期于我院体检的健康儿童（对照组）为研究对象,检测两组血清维生素A、维生素E水平、Ig水平,外周血T淋巴细胞亚群、NK细胞占比以及骨密度。分析维生素A、维生素E与Ig、T淋巴细胞亚群、NK细胞及骨密度的相关性。结果：感染组血清维生素A、维生素E、IgG、IgA、IgM及外周血CD3⁺T细胞百分比、CD4⁺T细胞百分比、CD3^-CD56⁺ NK细胞百分比、CD56brightNK细胞百分比、CD56dimNK细胞百分比、桡骨和胫骨骨密度均低于对照组（P＜0.05）,外周血CD8⁺T细胞百分比高于对照组（P＜0.05）。血清维生素A及维生素E水平与外周血CD8⁺T细胞百分比呈负相关（P＜0.05）,与IgG、IgA、IgM水平,外周血CD3⁺ T细胞百分比、CD4⁺T细胞百分比、CD3^-CD56⁺ NK细胞百分比、CD56brightNK细胞百分比、CD56dimNK细胞百分比、桡骨和胫骨骨密度呈正相关（P＜0.05）。结论：RRTI患儿血清维生素A、维生素E水平明显降低,且与免疫功能障碍和骨密度降低有关。     

Immune cell phenotype and functional defects in Netherton syndrome

Background

Netherton syndrome (NS) is a rare life-threatening syndrome caused by SPINK5 mutations leading to a skin barrier defect and a severe atopic diathesis. NS patients are prone to bacterial infections, but the understanding of the underlying immune deficiency is incomplete.

Results

We analyzed blood lymphocyte phenotypes and function in relation to clinical infections in 11 Finnish NS patients, aged 3 to 17?years, and healthy age-matched controls. The proportion of B cells (CD19⁺) and naïve B cells (CD27^?, IgD⁺) were high while memory B cells (CD27⁺) and switched memory B cells (CD27⁺IgM^?IgD^?), crucial for the secondary response to pathogens, was below or in the lowest quartile of the reference values in 8/11 (73%) and 9/11 (82%) patients, respectively. The proportion of activated non-differentiated B cells (CD21^low, CD38l^ow) was below or in the lowest quartile of the reference values in 10/11 (91%) patients. Despite normal T cell counts, the proportion of naïve CD4⁺ T cells was reduced significantly and the proportion of CD8⁺ T central memory significantly elevated. An increased proportion of CD57⁺ CD8⁺ T cells indicated increased differentiation potential of the T cells. The proportion of cytotoxic NK cells was elevated in NS patients in phenotypic analysis based on CD56DIM, CD16⁺ and CD27^? NK cells but in functional analysis, decreased expression of CD107a/b indicated impaired cytotoxicity.The T and NK cell phenotype seen in NS patients also significantly differed from that of age-matched atopic dermatitis (AD) patients, indicating a distinctive profile in NS. The frequency of skin infections correlated with the proportion of CD62L⁺ T cells, naïve CD4⁺ and CD27⁺ CD8⁺ T cells and with activated B cells. Clinically beneficial intravenous immunoglobulin therapy (IVIG) increased naïve T cells and terminal differentiated effector memory CD8⁺ cells and decreased the proportion of activated B cells and plasmablasts in three patients studied.

Conclusions

This study shows novel quantitative and functional aberrations in several lymphocyte subpopulations, which correlate with the frequency of infections in patients with Netherton syndrome. IVIG therapy normalized some dysbalancies and was clinically beneficial.

     

Altered effector function of peripheral cytotoxic cells in COPD

Background

There is mounting evidence that perforin and granzymes are important mediators in the lung destruction seen in COPD. We investigated the characteristics of the three main perforin and granzyme containing peripheral cells, namely CD8⁺ T lymphocytes, natural killer (NK; CD56⁺CD3^-) cells and NKT-like (CD56⁺CD3⁺) cells.

Methods

Peripheral blood mononuclear cells (PBMCs) were isolated and cell numbers and intracellular granzyme B and perforin were analysed by flow cytometry. Immunomagnetically selected CD8+ T lymphocytes, NK (CD56⁺CD3^-) and NKT-like (CD56⁺CD3⁺) cells were used in an LDH release assay to determine cytotoxicity and cytotoxic mechanisms were investigated by blocking perforin and granzyme B with relevant antibodies.

Results

The proportion of peripheral blood NKT-like (CD56⁺CD3⁺) cells in smokers with COPD (COPD subjects) was significantly lower (0.6%) than in healthy smokers (smokers) (2.8%, p < 0.001) and non-smoking healthy participants (HNS) (3.3%, p < 0.001). NK (CD56⁺CD3^-) cells from COPD subjects were significantly less cytotoxic than in smokers (16.8% vs 51.9% specific lysis, p < 0.001) as were NKT-like (CD56⁺CD3⁺) cells (16.7% vs 52.4% specific lysis, p < 0.001). Both cell types had lower proportions expressing both perforin and granzyme B. Blocking the action of perforin and granzyme B reduced the cytotoxic activity of NK (CD56⁺CD3^-) and NKT-like (CD56⁺CD3⁺) cells from smokers and HNS.

Conclusion

In this study, we show that the relative numbers of peripheral blood NK (CD56⁺CD3^-) and NKT-like (CD56⁺CD3⁺) cells in COPD subjects are reduced and that their cytotoxic effector function is defective.     

Granulocyte-macrophage colony-stimulating factor increases the proportion of circulating dendritic cells after autologous but not after allogeneic hematopoietic stem cell transplantation

Background aimsGranulocyte–macrophage (GM) colony-stimulating factor (CSF) has been used as an adjuvant in cancer immunotherapy. We tested the hypothesis that GM-CSF (Leukine^®; sargramostim) improves immune reconstitution after hematopoietic stem cell transplantation (HSCT) based on our prior in vitro work that demonstrated the pro-inflammatory effects of GM-CSF on dendritic cells (DC).MethodsGM-CSF was administered to donors, along with standard granulocyte (G) CSF, during stem cell mobilization, and to recipients from the day prior to transplant until engraftment. Eighteen patients consented to the GM-CSF⁺ protocol and were compared with 17 matched controls undergoing HSCT during the same time period (GM-CSF^?).ResultsNumbers of white blood cells (WBC) and CD34⁺ stem cells in the graft were comparable to controls. Surprisingly, contrary to our hypothesis, the allogeneic donor graft had significantly decreased numbers of CD3⁺ T cells and their subsets (CD4⁺, CD4⁺ CD45RA⁺, CD4⁺ CD45RO⁺, CD8⁺ and CD8⁺ CD45RO⁺), DC (both myeloid and plasmacytoid) and natural killer (NK) cells (CD16⁺ CD56⁺). In the GM-CSF arm, following allogeneic transplantation, the levels of DC, T cells and NK cells did not increase with treatment. Conversely, autologous transplant patients receiving GM-CSF had a higher proportion of DC at the time of engraftment.ConclusionsThese findings demonstrate that administration of GM-CSF improves DC reconstitution after autologous rather than allogeneic HSCT.     

培美曲塞化疗对非小细胞肺癌患者T淋巴细胞亚群的影响

目的:探究培美曲塞化疗对非小细胞肺癌患者T淋巴细胞亚群的影响,评价其临床应用价值。方法:选取2016年3月到2017年3月我院肿瘤内科收治的100例非小细胞肺癌患者进行研究,其中,50例非小细胞肺癌患者采用培美曲塞化疗、50例采用多西他赛进行化疗。比较患者治疗前后T淋巴细胞亚群变化情况。结果:与化疗前比较,非小细胞肺癌患者化疗后CD_3~+、CD_4~+、CD_4~+/CD_8~+比值以及NK细胞显著上升,而CD_8~+显著下降(P0.05)。为了验证培美曲塞对非小细胞肺癌患者的安全性,本研究将部分患者采用多西他赛化疗,非小细胞肺癌患者培美曲塞和多西他赛化疗后CD_3~+、CD_4~+、CD_4~+/CD_8~+比值以及NK细胞相比差异不大(P0.05)。此外,两组不良反应如恶心、呕吐,细胞减少,血小板减少,肾功能障碍差异不大(P0.05)。结论:培美曲塞化疗可以显著改变非小细胞患者的T淋巴细胞亚群构成,提高患者免疫功能和治疗效果。     

Participation of NK1.1+ T Cells in the Rejection of lpr αβT Cells When Bone Marrow Cells of Ipr Mice Are Transplanted into B6 Mice

When C57BL/6 (B6) mice were irradiated (9 Gy) and received bone marrow (BM) cells of B6-lpr/lpr mouse origin (i.e., lpr→B6), all mice died within 6 days. In the irradiated B6 mice, radioresistant CD3^? IL-2Rβ⁺ NK cells and IL-2Rβ CD3^int cells (i.e., CD3^int cells of extrathymic origin) remained, especially in the liver. There were two subsets, NK1.1⁺ and NK1.1^?, among the IL-2Rβ⁺ CD3^int cells. However, the NK1.1⁺ subset (i.e., NK1.1⁺ T cells) was much more radioresistant, and the majority of CD3^int cells belonged to this subset in irradiated mice. The expansion of lymphocytes from injected BM cells did not occur in the irradiated B6 mice. However, such expansion did take place in irradiated B6-lpr/lpr mice injected with both BM cells of B6-lpr/lpr and B6 origin. As a result, the mice subjected to BM cells survived. Irradiated B6 mice were treated in vivo with anti-NK1.1 mAb or anti-asialoGM₁ antibody to eliminate NK cells alone or both NK cells and NK1.1⁺ T cells. When irradiated B6 mice were pretreated with anti-NK1.1 mAb, the mice could survive. These results suggest that intact NK1.1⁺ T cells of extrathymic origin may recognize abnormal BM cells with the lpr gene and inhibit the expansion of lymphocytes, including abnormal double-negative CD4^?8^? cells, in B6-lpr/lpr mice. To inhibit the expansion of lymphocytes, mechanisms other than Fas ligand/Fas molecules on extrathymic T cells may be responsible.     

Immune reconstitution in patients with Fanconi anemia after allogeneic bone marrow transplantation

Background aimsFanconi anemia is an autosomal recessive or X-linked genetic disorder characterized by bone marrow (BM) failure/aplasia. Failure of hematopoiesis results in depletion of the BM stem cell reservoir, which leads to severe anemia, neutropenia and thrombocytopenia, frequently requiring therapeutic interventions, including hematopoietic stem cell transplantation (HSCT). Successful BM transplantation (BMT) requires reconstitution of normal immunity.MethodsIn the present study, we performed a detailed analysis of the distribution of peripheral blood subsets of T, B and natural killer (NK) lymphocytes in 23 patients with Fanconi anemia before and after BMT on days +30, +60, +100, +180, +270 and +360. In parallel, we evaluated the effect of related versus unrelated donor marrow as well as the presence of graft-versus-host disease (GVHD).ResultsAfter transplantation, we found different kinetics of recovery for the distinct major subsets of lymphocytes. NK cells were the first to recover, followed by cytotoxic CD8⁺ T cells and B cells, and finally CD4⁺ helper T cells. Early lymphocyte recovery was at the expense of memory cells, potentially derived from the graft, whereas recent thymic emigrant (CD31⁺ CD45RA⁺) and naive CD4⁺ or CD8⁺ T cells rose only at 6 months after HSCT, in the presence of immunosuppressive GVHD prophylactic agents. Only slight differences were observed in the early recovery of cytotoxic CD8⁺ T cells among those cases receiving a graft from a related donor versus an unrelated donor. Patients with GVHD displayed a markedly delayed recovery of NK cells and B cells as well as of regulatory T cells and both early thymic emigrant and total CD4⁺ T cells.ConclusionsOur results support the utility of post-transplant monitoring of a peripheral blood lymphocyte subset for improved follow-up of patients with Fanconi anemia undergoing BMT.     

地佐辛联合舒芬太尼对开胸患者术后镇痛镇静效果、血流动力学及免疫功能的影响

目的:探讨地佐辛联合舒芬太尼对开胸患者术后镇痛镇静效果、血流动力学及免疫功能的影响。方法:选取2017年1月~2018年12月期间新疆医科大学附属第一医院收治的拟行开胸手术患者70例,根据术后镇痛药物的不同将患者分为对照组(n=35)和研究组(n=35),对照组术后给予舒芬太尼镇痛,研究组术后给予地佐辛联合舒芬太尼镇痛。比较两组患者镇痛镇静效果、血流动力学、免疫功能及不良反应发生情况。结果:研究组术后6 h、12 h、24 h视觉疼痛模拟评分法(VAS)评分呈逐渐递减趋势,且低于对照组(P<0.05)。研究组术后3 h、6 h、12 h、24 h Ramsay镇静评分均低于对照组(P<0.05)。研究组术后5 min、3 h、12 h心率(HR)、舒张压(DBP)、收缩压(SBP)均低于对照组(P<0.05)。两组患者术后1 d、7 d CD3^+、CD4^+/CD8^+、自然杀伤(NK)细胞水平均呈先降低后升高趋势,术后7 d可恢复至术前水平(P<0.05),研究组术后1 d CD3^+、CD4^+/CD8^+、NK细胞水平高于对照组(P<0.05)。研究组不良反应发生率低于对照组(P<0.05)。结论:开胸患者术后应用地佐辛联合舒芬太尼,镇静镇痛效果较好,可有效维持患者血流动力学稳定,减轻机体免疫功能抑制,安全可靠。     

Changes in blood lymphocyte numbers with age in vivo and their association with the levels of cytokines/cytokine receptors

Background

Alterations in the number and composition of lymphocytes and their subsets in blood are considered a hallmark of immune system aging. However, it is unknown whether the rates of change of lymphocytes are stable or change with age, or whether the inter-individual variations of lymphocyte composition are stable over time or undergo different rates of change at different ages. Here, we report a longitudinal analysis of T- and B-cells and their subsets, and NK cells in the blood of 165 subjects aged from 24 to 90 years, with each subject assessed at baseline and an average of 5.6 years follow-up.

Results

The rates of change of T-(CD4⁺ and CD8⁺) and B-cells, and NK cells were relative stable throughout the adult life. A great degree of individual variations in numbers of lymphocytes and their subsets and in the rates of their changes with age was observed. Among them, CD4⁺ T cells exhibited the highest degree of individual variation followed by NK cells, CD8⁺ T cells, and B cells. Different types of lymphocytes had distinct trends in their rates of change which did not appear to be influenced by CMV infection. Finally, the rates of CD4⁺, CD8⁺ T cells, naive CD4⁺ and naïve CD8⁺ T cells were closely positively correlated.

Conclusion

Our findings provide evidence that the age-associated changes in circulating lymphocytes were at relative stable rates in vivo in a highly individualized manner and the levels of selected cytokines/cytokine receptors in serum might influence these age-associated changes of lymphocytes in circulation.

     

广泛性焦虑障碍患者人格特征与免疫功能、甲状腺激素和神经内分泌激素的相关性分析

摘要 目的：探讨广泛性焦虑障碍（GAD）患者人格特征与免疫功能、甲状腺激素和神经内分泌激素的相关性。方法：选择2019年1月~2020年12月北部战区空军医院心理科收治的GAD患者80例作为研究组,选择同期于北部战区空军医院体检的健康志愿者80例作为对照组。应用艾森克人格问卷简式量表中国版（EPQ-RSC）对受试者人格特征进行评价,比较两组EPQ-RSC评分结果、血液中CD3⁺、CD4⁺、自然杀伤细胞（NK）比例,血清白细胞介素（IL）-2、IL-6,三碘甲状腺原氨酸（T3）、甲状腺素（T4）、游离三碘甲状腺原氨酸（FT3）、游离甲状腺素（FT4）、促甲状腺激素（TSH）、去甲肾上腺素（NE）、促肾上腺皮质激素（ACTH）、皮质醇（CS）水平,并分析其相关性。结果：研究组精神质（P）、神经质（N）评分及EPQ-RSC总分均显著高于对照组（P<0.05）,研究组CD3⁺、CD4⁺、NK比例显著低于对照组,血清IL-2、IL-6水平显著高于对照组（P<0.05）。研究组血清FT3、FT4水平显著低于对照组（P<0.05）。研究组血清CS水平显著低于对照组,NE、ACTH水平显著高于对照组（P<0.05）。Pearson相关分析显示,GAD患者N、P评分及EPQ-RSC总分与CD3⁺、CD4⁺、NK、FT3、FT4、CS呈负相关（P<0.05）,与IL-2、IL-6、NE、ACTH呈正相关（P<0.05）。结论：GAD患者存在免疫功能损伤、神经内分泌激素和甲状腺激素水平异常,且均与患者P、N倾向的人格特征有关。     

Perforin, granzyme B, and FasL expression by peripheral blood T lymphocytes in emphysema

Background

It is generally accepted that emphysematous lungs are characterized by an increase in the numbers of neutrophils, macrophages, and CD8⁺ T lymphocytes, the lasts having increased cytotoxic activity. Because systemic inflammation is also a component of emphysema, we hypothesize that peripheral CD8⁺ T lymphocytes of emphysematous smokers who show evidence of systemic inflammation will have higher expression of cytotoxic molecules.

Methods

We assessed parameters of systemic inflammation in normal individuals (smokers or non-smokers) and in emphysematous subjects with an active smoking history by measuring serum interleukine-6, C-reactive protein, and tumor necrosis factor. Expression of perforin, granzyme B, and FasL protein by CD8⁺ T lymphocytes, CD4⁺ T lymphocytes, and natural killer cells were assessed by flow cytometry while perforin, granzyme B, and FasL mRNA expression were measured on purified systemic CD8⁺ T lymphocytes by real-time PCR.

Results

Emphysematous smokers had higher levels of serum interleukine-6 than normal subjects. Even with the presence of systemic inflammation in emphysematous smokers, the percentage of peripheral CD8⁺ T lymphocytes, CD4⁺ T lymphocytes, and NK cells expressing perforin and granzyme B protein was not different between the three groups.

Conclusion

Despite evidence of systemic inflammation, peripheral T lymphocytes of emphysematous smokers did not show higher levels of cytotoxic markers, suggesting that increase of activated T lymphocytes in the emphysematous lung may be due to either activation in the lung or specific peripheral recruitment.     

Molecular targeting of hepatocyte growth factor by an antagonist,NK4, in the treatment of rheumatoid arthritis

Introduction

Hepatocyte growth factor (HGF) is a potent proangiogenic molecule that induces neovascularization. The HGF antagonist, NK4, competitively antagonizes HGF binding to its receptor. In the present study, we determined the inhibitory effect of NK4 in a rheumatoid arthritis (RA) model using SKG mice.

Methods

Arthritis was induced in SKG mice by a single intraperitoneal injection of β-glucan. Recombinant adenovirus containing NK4 cDNA (AdCMV.NK4) was also injected intravenously at the time of or 1 month after β-glucan injection. Ankle bone destruction was examined radiographically. The histopathologic features of joints were examined using hematoxylin and eosin and immunohistochemical staining. Enzyme-linked immunosorbent assays were used to determine the serum levels of HGF, interferon γ (IFN-γ, interleukin 4 (IL-4) and IL-17 production by CD4⁺ T cells stimulated with allogeneic spleen cells.

Results

The intravenous injection of AdCMV.NK4 into SKG mice suppressed the progression of β-glucan-induced arthritis. Bone destruction was also inhibited by NK4 treatment. The histopathologic findings of the ankles revealed that angiogenesis, inflammatory cytokines and RANKL expression in synovial tissues were significantly inhibited by NK4 treatment. Recombinant NK4 (rNK4) proteins inhibited IFN-γ, IL-4 and IL-17 production by CD4⁺ T cells stimulated with allogeneic spleen cells.

Conclusions

These results indicate that NK4 inhibits arthritis by inhibition of angiogenesis and inflammatory cytokine production by CD4⁺ T cells. Therefore, molecular targeting of angiogenic inducers by NK4 can potentially be used as a novel therapeutic approach for the treatment of RA.     

Expression of murine killer immunoglobulin-like receptor KIRL1 on CD1d-independent NK1.1<Superscript>+</Superscript> T cells

Three mouse killer immunoglobulin-like receptors (KIRs), namely, KIR3DL1, KIRL1, and KIRL2, have recently been identified in C56BL/6 (B6) mice. However, only two Kir genes are found in the B6 mouse genome sequence data base. To clarify this discrepancy, we cloned Kir cDNAs from multiple strains of mice. Sequencing of the cDNA clones showed that the Kir3dl1 gene is found in C3H/HeJ and CBA/J but not in B6 mice. Analysis of the single nucleotide polymorphism data base suggested that Kir3dl1 is the C3H/HeJ and CBA/J allele of Kirl1. We generated mAb to the recombinant KIRL1 protein to investigate its expression pattern. The anti-KIRL1 mAb bound to NK1.1⁺ T cells but only very weakly or at undetectable levels to other lymphocytes including natural killer (NK) cells and conventional T cells. Among NK1.1⁺ T cells, conventional NK T cells stained with CD1d tetramer did not significantly bind anti-KIRL1 mAb, whereas CD1d-tetramer-negative subset was KIRL1-positive. Furthermore, the expression of KIRL1 is readily detected on NK1.1⁺ T cells from β₂-microglobulin-deficient B6 mice. Thus, KIRL1 is predominantly expressed on CD1d-independent NK1.1⁺ T cells.     

腹腔镜全结肠系膜切除术治疗老年结肠癌患者的近期疗效评价及对机体免疫力的影响

目的:研究腹腔镜全结肠系膜切除术对老年结肠癌患者的近期疗效评价及机体免疫力的影响,为临床治疗结肠癌提供依据。方法:选择2014年6月至2016年6月我院收治的136例老年结肠癌患者为研究对象,按照患者治疗意愿分为两组。治疗组57例行腹腔镜下全结肠系膜切除术,对照组79例行开腹手术。比较两组患者的手术时间、淋巴结清扫数量、术中出血量、肛门排气时间、下床活动时间、术后住院时间及术后并发症发生率。于术前1 d、术后1 d及术后3 d采用流式细胞仪检测两组患者外周血中T淋巴细胞亚群(CD3~+、CD4~+、CD8~+)及自然杀伤(NK)细胞;于术前1 d、术后1 d及术后3 d采用酶联免疫吸附法(ELISA)测定两组患者血清C反应蛋白(CRP)、可溶性白细胞介素-2受体(sIL-2R)、白细胞介素-6(IL-6)、白细胞介素-8(IL-8)浓度。结果:与对照组比较,治疗组术中出血量减少,肛门排气时间、下床活动时间及术后住院时间均缩短,差异均有统计学意义(P0.05)。两组患者淋巴结清扫数量及手术时间比较,差异无统计学意义(P0.05)。治疗组并发症发生率为7.02%,低于对照组的17.72%,差异具有统计学意义(P0.05)。治疗组患者术后1 d和术后3 d外周血中T细胞亚群CD3~+、CD4~+和NK细胞所占比例及CD4~+/CD8~+的比值与术前1 d相比有所减小(P0.05),且术后1 d的CD4~+/CD8~+的比值和NK细胞低于术后3 d(P0.05);两组术后1 d及术后3 d外周血中CD8~+所占比例与术前1 d有所升高,但是差异无统计学意义(P0.05),且术后1 d的CD8~+所占比例与术后3d比较无明显差异(P0.05)。治疗组患者术后1 d和术后3天外周血中CD3~+、CD4~+与对照组比较,差异无统计学意义(P0.05),CD4~+/CD8~+的比值和NK细胞高于对照组,差异有统计学意义(P0.05)。治疗组术后1 d及术后3 d血清CRP、sIL-2R、IL-6、IL-8水平与均较术前1 d增高,术后3 d治疗组血清中CRP、sIL-2R、IL-6、IL-8水平低于术后1 d(P0.05),治疗组术后1 d及术后3 d血清中CRP、sIL-2R、IL-6、IL-8水平均低于对照组,差异均具有统计学意义(P0.05)。结论:腹腔镜全结肠系膜切除术与开腹手术相比,近期疗效更好,具有创口小、出血量少、术后恢复快、术后并发症发生率低及对机体免疫功能影响小等优点,可应用于老年结肠癌的治疗。     

A combination of Olea europaea leaf extract and Spirodela polyrhiza extract alleviates atopic dermatitis by modulating immune balance and skin barrier function in a 1-chloro-2,4-dinitrobenzene-induced murine model

BackgroundAtopic dermatitis is a chronic inflammatory skin disease in humans. Although Olea europaea leaf extract (OLE) and Spirodela polyrhiza extract (SPE) have been used to protect against skin damage, the effects of their combined administration on atopic dermatitis have yet to studied.PurposeIn this study, we evaluated the potential therapeutic effects of an OLE and SPE combination on the progression of atopic dermatitis and the possible mechanisms underlying these effects in 1-chloro-2,4-dinitrobenzene (DNCB)-treated NC/Nga mice.MethodsAtopic dermatitis was induced by topical application of 0.2% w/v DNCB prepared in an olive oil:acetone solution (1:3), and thereafter OLE, SPE and OLE + SPE were administered orally for 5 weeks. We determined atopic dermatitis symptoms, serum IgE levels, and levels of cytokine- and gene expression in the dorsal skin and splenocytes, and performed histological and immune cell subtype analyses. The expression of skin barrier-related proteins (filaggrin, sirtuin 1, and claudin 1) was also evaluated.ResultsThe OLE + SPE combination significantly ameliorated atopic dermatitis symptoms, including dermatitis scores, and reduced epidermal thickness and infiltration of different inflammatory cells in mice with DNCB-induced atopic dermatitis. It also significantly reduced the number of CD4⁺, CD8⁺, and CD4⁺/CD69⁺ T cells; immunoglobulin E-producing B cells (CD23⁺/B220⁺) in the axillary lymph nodes; CD3⁺ T-cell eosinophils (chemokine–chemokine receptor 3⁺/CD11b⁺) in the skin; and CD3⁺ T cells, immunoglobulin E-producing B cells (CD23⁺/B220⁺), and eosinophils in peripheral blood mononuclear cells. Additionally, the experimental combination lowered levels of serum immunoglobulin E and histamine, as well as Th2-mediated cytokines, and interleukin-4, -5, and -13, whereas it increased the levels of Th1-mediated cytokine interferon-γ in splenocytes. Furthermore, the preparation significantly restored expression of the skin barrier-related proteins filaggrin, sirtuin 1, and claudin 1, and also reduced the expression of the inflammatory cytokine interleukin-6 and chemokine–chemokine receptor 3, as well as the pruritus-related cytokine interleukin-31 and interleukin-31 receptor, in atopic dermatitis skin lesions.ConclusionTaken together, our findings indicate that administration of a combination of OLE and SPE can alleviate atopic dermatitis symptoms by regulating immune balance and skin barrier function and may be an effective therapeutic option for the treatment of atopic dermatitis.     

A proportion of CD4+ T cells from patients with chronic Chagas disease undergo a dysfunctional process,which is partially reversed by benznidazole treatment

BackgroundSigns of senescence and the late stages of differentiation associated with the more severe forms of Chagas disease have been described in the Trypanosoma cruzi antigen-specific CD4⁺ T-cell population. However, the mechanisms involved in these functions are not fully known. To date, little is known about the possible impact of benznidazole treatment on the T. cruzi-specific functional response of CD4⁺ T cells.Methodology/Principal findingsThe functional capacity of CD4⁺ T cells was analyzed by cytometric assays in chronic Chagas disease patients, with indeterminate form (IND) and cardiac alterations (CCC) (25 and 15, respectively) before and after benznidazole treatment. An increase in the multifunctional capacity (expression of IFN-γ, IL-2, TNF-α, perforin and/or granzyme B) of the antigen-specific CD4⁺ T cells was observed in indeterminate versus cardiac patients, which was associated with the reduced coexpression of inhibitory receptors (2B4, CD160, CTLA-4, PD-1 and/or TIM-3). The functional profile of these cells shows statistically significant differences between IND and CCC (p<0.001), with a higher proportion of CD4⁺ T cells coexpressing 2 and 3 molecules in IND (54.4% versus 23.1% and 4.1% versus 2.4%, respectively). A significant decrease in the frequencies of CD4⁺ T cells that coexpress 2, 3 and 4 inhibitory receptors was observed in IND after 24–48 months of treatment (p<0.05, p<0.01 and p<0.05, respectively), which was associated with an increase in antigen-specific multifunctional activity. The IND group showed, at 9–12 months after treatment, an increase in the CD4⁺ T cell subset coproducing three molecules, which were mainly granzyme B⁺, perforin⁺ and IFN-γ⁺ (1.4% versus 4.5%).Conclusions/SignificanceA CD4⁺ T cell dysfunctional process was detected in chronic Chagas disease patients, being more exacerbated in those patients with cardiac symptoms. After short-term benznidazole treatment (9–12 months), indeterminate patients showed a significant increase in the frequency of multifunctional antigen-specific CD4⁺ T cells.     

IL-15 Superagonist Expands mCD8+ T,NK and NKT Cells after Burn Injury but Fails to Improve Outcome during Burn Wound Infection

Background

Severely burned patients are highly susceptible to opportunistic infections and sepsis, owing to the loss of the protective skin barrier and immunological dysfunction. Interleukin-15 (IL-15) belongs to the IL-2 family of common gamma chain cytokines and stimulates the proliferation and activation of T (specifically memory CD8), NK and NKT cells. It has been shown to preserve T cell function and improve survival during cecal ligation and puncture (CLP)-induced sepsis in mice. However, the therapeutic efficacy of IL-15 or IL-15 superagonist (SA) during infection after burn injury has not been evaluated. Moreover, very few, if any, studies have examined, in detail, the effect of burn injury and infection on the adaptive immune system. Thus, we examined the effect of burn and sepsis on adaptive immune cell populations and the effect of IL-15 SA treatment on the host response to infection.

Methods

Mice were subjected to a 35% total body surface area burn, followed by wound infection with Pseudomonas aeruginosa. In some experiments, IL-15 SA was administered after burn injury, but before infection. Leukocytes in spleen, liver and peritoneal cavity were characterized using flow cytometry. Bacterial clearance, organ injury and survival were also assessed.

Results

Burn wound infection led to a significant decline in total white blood cell and lymphocyte counts and induced organ injury and sepsis. Burn injury caused decline in CD4⁺ and CD8⁺ T cells in the spleen, which was worsened by infection. IL-15 treatment inhibited this decline and significantly increased cell numbers and activation, as determined by CD69 expression, of CD4⁺, CD8⁺, B, NK and NKT cells in the spleen and liver after burn injury. However, IL-15 SA treatment failed to prevent burn wound sepsis-induced loss of CD4⁺, CD8⁺, B, NK and NKT cells and failed to improve bacterial clearance and survival.

Conclusion

Cutaneous burn injury and infection cause significant adaptive immune dysfunction. IL-15 SA does not augment host resistance to burn wound sepsis in mice despite inducing proliferation and activation of lymphocyte subsets.     

Substantial increase in the frequency of circulating CD4+NKG2D+ T cells in patients with cervical intraepithelial neoplasia grade 1

Background

The NKG2D receptor confers important activating signals to NK cells via ligands expressed during cellular stress and viral infection. This receptor has generated great interest because not only is it expressed on NK cells, but it is also seen in virtually all CD8⁺ cytotoxic T cells and is classically considered absent in CD4⁺ T cells. However, recent studies have identified a distinctive population of CD4⁺ T cells that do express NKG2D, which could represent a particular cytotoxic effector population involved in viral infections and chronic diseases. On the other hand, increased incidence of human papillomavirus-associated lesions in CD4⁺ T cell-immunocompromised individuals suggests that CD4⁺ T cells play a key role in controlling the viral infection. Therefore, this study was focused on identifying the frequency of NKG2D-expressing CD4⁺ T cells in patients with cervical intraepithelial neoplasia (CIN) 1. Additionally, factors influencing CD4⁺NKG2D⁺ T cell expansion were also measured.

Results

Close to 50% of patients with CIN 1 contained at least one of the 37 HPV types detected by our genotyping system. A tendency for increased CD4⁺ T cells and CD8⁺ T cells and decreased NK cells was found in CIN 1 patients. The percentage of circulating CD4⁺ T cells co-expressing the NKG2D receptor significantly increased in women with CIN 1 versus control group. Interestingly, the increase of CD4⁺NKG2D⁺ T cells was seen in patients with CIN 1, despite the overall levels of CD4⁺ T cells did not significantly increase. We also found a significant increase of soluble MICB in CIN 1 patients; however, no correlation with the presence of CD4⁺NKG2D⁺ T cells was seen. While TGF-beta was significantly decreased in the group of CIN 1 patients, both TNF-alpha and IL-15 showed a tendency to increase in this group.

Conclusions

Taken together, our results suggest that the significant increase within the CD4⁺NKG2D⁺ T cell population in CIN 1 patients might be the result of a chronic exposure to viral and/or pro-inflammatory factors, and concomitantly might also influence the clearance of CIN 1-type lesion.     

卡瑞利珠单抗联合卡培他滨用于一线治疗复发转移鼻咽癌患者维持治疗疗效及对患者免疫功能的影响

摘要 目的：探讨卡瑞利珠单抗联合卡培他滨用于一线治疗复发转移鼻咽癌（NPC）患者维持治疗的疗效及对患者免疫功能的影响。方法：选取2020年1月至2022年1月钦州市第一人民医院收治的80例复发转移NPC患者,按照随机数字表法分为对照组和观察组,每组各40例。两组均接受吉西他滨联合顺铂化疗,对照组化疗后接受卡瑞利珠单抗单药维持治疗至1年,观察组化疗后接受卡瑞利珠单抗联合卡培他滨维持治疗至1年。比较两组疗效,不良反应,治疗前后B淋巴细胞亚群和自然杀伤（NK）细胞所占百分比的差异。结果：观察组客观缓解率（ORR）、疾病控制率（DCR）高于对照组（P＜0.05）。两组治疗后外周血CD5⁺B细胞、CD5⁺CD19⁺ B细胞、CD3^-CD56⁺NK细胞占比降低（P＜0.05）,但观察组治疗后外周血CD5⁺B细胞、CD5⁺CD19⁺ B细胞、CD3^-CD56⁺NK细胞占比高于对照组（P＜0.05）。两组Ⅲ度以上不良反应发生率比较差异无统计学意义（P＞0.05）。结论：卡瑞利珠单抗联合卡培他滨一线维持治疗复发转移NPC可提高临床疗效,减轻对细胞免疫功能的影响,且安全可靠。