丙型肝炎病毒感染的细胞免疫研究进展

本文简述近几年来丙型肝炎病毒（ＨＣＶ）感染与宿主细胞免疫应答之间关系的研究进展,重点对ＨＣＶ感染后体内ＣＤ４＾＋Ｔ细胞、ＣＤ８＾＋Ｔ细胞和细胞毒性Ｔ淋巴细胞（ＣＴＬ）的免疫应答及其在免疫致病中的作用进行了讨论。     

双歧杆菌活菌制剂对慢性乙肝病人细胞免疫功能影响的观察

在应用双歧杆菌活菌制剂治疗慢乙肝期间,重点观察了Ｔ细胞亚群（ＣＤ３,ＣＤ４,ＣＤ８）、ＮＫ细胞（ＣＤ（１６））、白细胞介素Ⅱ（ＩＬ－２）分泌细胞、肿瘤坏死因子（ＴＮＦ）等细胞免疫指标治疗前后的动态变化,同时观察了病人血内毒素水平的动态变化和乙肝病毒标志物（ＨＢＶＭ）的改变。结果表明：（１）与对照组比较,双歧杆菌活菌制剂可使慢乙肝病人ＣＤ３＋,ＣＤ４＋数目明显增多,而对ＣＤ８＋细胞数目无明显影响;（２）双歧杆菌活菌制剂可使ＣＡＨ组的ＣＤ１６＋细胞显著增多（ｐ＜０．０５）;使ＣＡＨ组和ＣＰＨ组的ＩＬ－２分泌细胞均有非常显著和显著增加（分别ｐ＜０．０１和ｐ＜０．０５）;（３）ＣＡＨ组病人血中内毒素和ＴＮＦ水平在双歧杆菌活菌制剂治疗后,匀出现非常显著降低（ｐ＜０．０１）;ＣＰＨ组ＴＮＦ水平较对照组无显著变化,但内毒素水平较对照组显著降低（ｐ＜０．０５）;（４）满疗程后（６０天）ＣＡＨ组有６例,ＣＰＨ组有５例ＨＢｅＡｇ阴转（分别为２６．０６％和２５．０％）,而对照组仅２例阴转（１３．３３％）,两治疗组与对照组比较有显著性差异（ｐ＜０．０５）。     

人类免疫缺陷病毒与程序性T细胞死亡

关于人类免疫缺陷病毒（ＨＩＶ）感染引发ＡＩＤＳ的发病机制尚不明确,本文介绍了一种新的致病机制,认为ＨＩＶ感染机体后可能诱导ＣＤ４＾＋和ＣＤ８＾＋Ｔ细胞发生程序性死亡,从而导致ＡＩＤＳ产生,并依此机制探讨有关抗ＨＩＶ感染的新途径。     

丙型肝炎病毒E2基因DNA免疫实验动物研究

将编码丙型肝炎病毒（ＨＣＶ）Ｅ２蛋白４１７～７５０位氨基酸的ＤＮＡ片段　克隆到真核表达载体ｐｃＤＮＡ　３．１（－）中的ＣＭＶ　ＩＥ启动子下游,构建成ＨＣＶ　Ｅ２重组真核表达质粒　ｐｃＥ２。ＥＬＩＳＡ法检测ｐｃＥ２　ＤＮＡ免疫兔血清中的Ｅ２抗体变化和维持规律,结果显示免疫２０ｄ已有　抗体产生,３０ｄ后开始进入高峰,４０ｄ时达到最高值,至第９０ｄ抗体水平保持平稳,抗体滴度　达到１∶１６００左右。流式细胞计数仪（ＦＡＣＳ）检测ｐｃＥ２　ＤＮＡ免疫鼠ＣＤ４＋、ＣＤ８＋Ｔ淋巴细胞变　化情况,与注射空载体ｐＣＤＮＡ３．１（－）的阴性鼠相比,ＣＤ４＋淋巴细胞水平略有上升,ＣＤ８＋　细胞水平有较大升高,增幅达３５．４６％。免疫组化检测结果显示注射ｐｃＥ２的小鼠组织中有明显　的阳性着色,而注射ｐｃＤＮＡ３．１（－）的对照组小鼠免疫组化结果为阴性。以上结果表明：ｐｃＥ２　在实验动物内表达出的ＨＣＶ　Ｅ２蛋白可以引起免疫动物的体液免疫应答和细胞免疫应答,尤其　是ＭＨＣ－１限制性杀伤性ＣＤ８＋Ｔ淋巴细胞水平的提高对清除　病毒是十分有利的,因此ＨＣＶ　Ｅ２　ＤＮＡ免疫有可能成为预防和治疗ＨＣＶ感染的一条新途径。     

CD28共刺激可促HIV阳性患者CD_4~ T细胞增殖

ＣＤ２８共刺激可促ＨＩＶ阳性患者ＣＤ＋４Ｔ细胞增殖ＣＤ＋４Ｔ细胞是人免疫缺陷病毒（ＨＩＶ）感染的主要细胞。由于在体外难以诱导ＨＩＶ感染者的自体ＣＤ＋４Ｔ细胞增殖,因此限制了许多疗法如基因治疗、免疫重建疗法的应用。已知在Ｔ细胞活化中,Ｔ细胞受体（ＴＣＲ...     

鸡传染性支气管炎病毒S1基因在昆虫细胞中表达水平初探

将含有鸡传染性支气管炎病毒 Ｓ１ 基因ｃ Ｄ Ｎ Ａ 的重组转移质粒ｐ Ｓ Ｘ Ｉ Ｖ Ｖ Ｉ＋ Ｘ３ Ｓ１ ． Ｈｏｌｔｅ 和ｐ Ｓ Ｘ Ｉ Ｖ Ｖ Ｉ＋ Ｘ３／４ Ｓ１ ． Ｈｏｌｔｅ 分别与粉纹夜蛾核型多角体病毒 Ｔｎ Ｎ Ｐ Ｖ Ｓ Ｖ Ｉ－ Ｇ Ｄ Ｎ Ａ（ Ｏ Ｃ Ｃ－ ,ｇａｌ＋ ） 共转染草地夜蛾（ Ｓｆ９） 细胞,经空斑纯化得到重组病毒 Ｔｎ Ｎ Ｐ Ｖ（ Ｘ３） Ｓ１ ． Ｈｏｌｔｅ Ｏ Ｃ Ｃ＋ 和 Ｔｎ Ｎ Ｐ Ｖ（ Ｘ３／４） Ｓ１ ． Ｈｏｌｔｅ Ｏ Ｃ Ｃ＋ 。将重组毒株分别感染 Ｔｎ５ Ｂ１ 细胞,并进行 Ｓ Ｄ Ｓ Ｐ Ａ Ｇ Ｅ 与 Ｗｅｓｔｅｒｎｂｌｏｔ 检测。结果表明, Ｔｎ Ｎ Ｐ Ｖ（ Ｘ３／４） Ｓ１ ． Ｈｏｌｔｅ Ｏ Ｃ Ｃ＋ 在感染的细胞中高效表达了 Ｓ１ 蛋白, Ｓ Ｄ Ｓ Ｐ Ａ Ｇ Ｅ 凝胶薄层色谱分析结果显示,感染病毒后７２ ｈ Ｓ１ 蛋白的表达量占细胞内总蛋白量的３５ ．８ ％ ,而 Ｔｎ Ｎ Ｐ Ｖ（ Ｘ３） Ｓ１ ． Ｈｏｌｔｅ Ｏ Ｃ Ｃ＋ 感染的细胞内检测不出 Ｓ１ 蛋白。经分析认为这一差异主要来自 Ｓ１ 基因翻译起始位点及其附近的周围环境。     

HSV—TK／GCV系统杀伤肝癌细胞的在体研究

目的和方法：将可产生含有ＨＳＶＴＫ基因逆转录病毒的包装细胞与ＢＥＬ７４０２肝癌细胞混合后,接种于裸鼠皮下,复制肝癌模型。确定ＨＳＶＴＫ基因转移后,给予动物注射ＧＣＶ,观察ＨＳＶＴＫ／ＧＣＶ系统基因治疗实验性肝癌的在体疗效。结果：在体条件下,逆转录病毒可将ＨＳＶＴＫ基因部分转导至肝癌细胞;ＧＣＶ作用后,ＴＫ＋中瘤细胞的生长受到明显抑制;电镜观察发现,ＧＣＶ杀伤ＴＫ＋肿瘤细胞的作用主要体现在细胞核;虽然组化染色分析表明ＴＫ基因转移效率只有１０％～３０％,但测量肿瘤体积显示ＨＳＶＴＫ／ＧＣＶ系统杀伤肝癌细胞的效果依然明显,故其作用机制可与“旁观者效应”有关。结论：ＨＳＶＴＫ／ＧＣＶ“自杀”基因系统有可能成为基因治疗肝癌的有效方法     

二甲亚砜与维甲酸对人卵巢癌细胞COC1的生长抑制及转化生长因子β_1表达的影响

本实验以人卵巢癌细胞株（ＣＯＣ１）为模型,观察诱导分化剂二甲亚砜（ＤＭＳＯ）与维甲酸（ＲＡ）对该细胞生长增殖、ＤＮＡ合成和转化生长因子β１（ＴＧＦβ１）在细胞内表达的影响。结果显示：ＤＭＳＯ与ＲＡ对人卵巢细胞ＣＯＣ１生长有明显的抑制作用,生长曲线表明作用５天后其生长抑制率分别为６２．７％和４２．１％;３Ｈ－胸腺嘧啶核苷（３Ｈ－ＴｄＲ）掺入实验说明ＤＭＳＯ组与ＲＡ组的单位时间计数率（ＣＰＭ）明显低于对照组（Ｐ＜０．０１）。用药３天后百分掺入抑制率分别为６０．４％与３７．９％,表明ＤＭＳＯ与ＲＡ抑制ＣＯＣ１细胞的ＤＮＡ合成;免疫细胞化学反应表明,ＤＭＳＯ或ＲＡ处理５天后,对照组细胞ＴＧＦβ１表达为阳性,定位于胞浆,而处理组细胞ＴＧＦβ１呈阴性或弱阳性反应。以上结果提示ＤＭＳＯ和ＲＡ对人卵巢癌细胞有一定的诱导分化作用。     

HBsAg携带者重叠感染HCV、HDV的血清学调查

对３６２名无症状高滴度ＨＢｓＡｇ携带者用ＲＩＡ法检测ＨＢｅＡｇ、Ａｎｔｉ－ＨＢｅ、Ａｎｔｉ－ＨＣＶ、Ａｎｔｉ－ＨＤＶ。结果表明,ＨＢｅＡｇ阳性率随ＨＢｓＡｇ滴度的增高而增加,且有显著性差异（Ｐ＜０．０５）。重叠感染以ＨＢＶ＋ＨＣＶ最高,占２７．６％;其次是ＨＢＶ＋ＨＤＶ,占９．１％;ＨＢＶ＋ＨＣＶ＋ＨＤＶ最少,占６．４％。但是,以上重叠感染率均与ＨＢｓＡｇ滴度及／或ＨＢｅＡｇ阳性率高低无关（Ｐ＞０．０５）。调查显示,无症状ＨＢｓＡｇ携带者中ＨＢＶ＋ＨＣＶ,或ＨＢＶ＋ＨＤＶ,或ＨＢＶ＋ＨＣＶ＋ＨＤＶ的重叠感染均可发生。     

人血树突状细胞对LAK细胞抗H_（7402）细胞影响的形态学观察

本研究结合光镜免疫组化和组织化学方法,在光镜和电镜下观察了人外周血树突状细胞（ＤＣ）对ＬＡＫ细胞杀伤人肝癌Ｈ７４０２细胞的影响。结果显示,（１）ＤＣ不仅能与ＬＡＩＣ细胞形成花环,还能与ＬＡＫ、Ｈ（７４０２）细胞相互接触形成细胞簇,常见呈明显坏死状态的Ｈ（７４０２）周围有ＤＣ和ＬＡＫ细胞围绕;（２）与Ｈ（７４０２）＋ＬＡＫ细胞组相比,在Ｈ（７４０２）＋ＬＡＫ＋ＤＣ组中,肿瘤细胞的坏死程度明显加重。提示人外周血ＤＣ在细胞免疫抗肿瘤过程中起重要作用。     

Antibody-independent antiviral function of memory CD4+ T cells in vivo requires regulatory signals from CD8+ effector T cells

Previous studies have shown that vaccine-primed CD4(+) T cells can mediate accelerated clearance of respiratory virus infection. However, the relative contributions of Ab and CD8(+) T cells, and the mechanism of viral clearance, are poorly understood. Here we show that control of a Sendai virus infection by primed CD4(+) T cells is mediated through the production of IFN-gamma and does not depend on Ab. This effect is critically dependent on CD8(+) cells for the expansion of CD4(+) T cells in the lymph nodes and the recruitment of memory CD4(+) T cells to the lungs. Passive transfer of a CD8(+) T cell supernatant into CD8(+) T cell-depleted, hemagglutinin-neuraminidase (HN)(421-436)-immune muMT mice substantially restored the virus-specific memory CD4(+) response and enhanced viral control in the lung. Together, the data demonstrate for the first time that in vivo primed CD4(+) T cells have the capacity to control a respiratory virus infection in the lung by an Ab-independent mechanism, provided that CD8(+) T cell "help" in the form of soluble factor(s) is available during the virus infection. These studies highlight the importance of synergistic interactions between CD4(+) and CD8(+) T cell subsets in the generation of optimal antiviral immunity.     

Aged Mice Exhibit a Severely Diminished CD8 T Cell Response following Respiratory Syncytial Virus Infection

Respiratory virus infections in the elderly result in increased rates of hospitalization and death. Respiratory syncytial virus (RSV) is a leading cause of severe virus-induced respiratory disease in individuals over the age of 65. CD8 T cells play a critical role in mediating RSV clearance. While it is clear that T cell immunity declines with age, it is not clear to what extent the CD8 T cell response to RSV is altered. Using aged BALB/c mice, we demonstrated that RSV-specific CD8 T cell responses were significantly reduced in the lungs of aged mice at the peak of the T cell response and that this decrease correlated with delayed viral clearance. Despite a decrease in the overall numbers of RSV-specific CD8 T cells during acute infection, their capacity to produce effector cytokines was not impaired. Following viral clearance, the RSV-specific memory CD8 T cells were similar in total number and phenotype in young and aged mice. Furthermore, following infection with a heterologous pathogen expressing an RSV epitope, RSV-specific memory CD8 T cells exhibited similar activation and ability to provide early control of the infection in young and aged mice. These data demonstrate a decrease in the capacity of aged mice to induce a high-magnitude acute CD8 T cell response, leading to prolonged viral replication, which may contribute to the increased disease severity of RSV infection observed for aged individuals.     

Potential of equine herpesvirus 1 as a vector for immunization

Key problems using viral vectors for vaccination and gene therapy are antivector immunity, low transduction efficiencies, acute toxicity, and limited capacity to package foreign genetic information. It could be demonstrated that animal and human cells were efficiently transduced with equine herpesvirus 1 (EHV-1) reconstituted from viral DNA maintained and manipulated in Escherichia coli. Between 13 and 23% of primary human CD3+, CD4+, CD8+, CD11b+, and CD19+ cells and more than 70% of CD4+ MT4 cells or various human tumor cell lines (MeWo, Huh7, HeLa, 293T, or H1299) could be transduced with one infectious unit of EHV-1 per cell. After intranasal instillation of EHV-1 into mice, efficient transgene expression in lungs was detectable. Successful immunization using EHV-1 was shown after delivery of the human immunodeficiency virus type 1 Pr55gag precursor by the induction of a Gag-specific CD8+ immune response in mice. Because EHV-1 was not neutralized by human sera containing high titers of antibodies directed against human herpesviruses 1 to 5, it is concluded that this animal herpesvirus has enormous potential as a vaccine vector, because it is able to efficiently transduce a variety of animal and human cells, has high DNA packaging capacity, and can conveniently be maintained and manipulated in prokaryotic cells.     

Susceptibility to acute mouse adenovirus type 1 respiratory infection and establishment of protective immunity in neonatal mice

There is an incomplete understanding of the differences between neonatal immune responses that contribute to the increased susceptibility of neonates to some viral infections. We tested the hypothesis that neonates are more susceptible than adults to mouse adenovirus type 1 (MAV-1) respiratory infection and are impaired in the ability to generate a protective immune response against a second infection. Following intranasal infection, lung viral loads were greater in neonates than in adults during the acute phase but the virus was cleared from the lungs of neonates as efficiently as it was from adult lungs. Lung gamma interferon (IFN-γ) responses were blunted and delayed in neonates, and lung viral loads were higher in adult IFN-γ(-/-) mice than in IFN-γ(+/+) controls. However, administration of recombinant IFN-γ to neonates had no effect on lung viral loads. Recruitment of inflammatory cells to the airways was impaired in neonates. CD4 and CD8 T cell responses were similar in the lungs of neonates and adults, although a transient increase in regulatory T cells occurred only in the lungs of infected neonates. Infection of neonates led to protection against reinfection later in life that was associated with increased effector memory CD8 T cells in the lungs. We conclude that neonates are more susceptible than adults to acute MAV-1 respiratory infection but are capable of generating protective immune responses.     

Murine model of interstitial cytomegalovirus pneumonia in syngeneic bone marrow transplantation: persistence of protective pulmonary CD8-T-cell infiltrates after clearance of acute infection

Interstitial pneumonia (IP) is a severe organ manifestation of cytomegalovirus (CMV) disease in the immunocompromised host, in particular in recipients of bone marrow transplantation (BMT). Diagnostic criteria for the definition of CMV-IP include clinical evidence of pneumonia together with CMV detected in bronchoalveolar lavage or lung biopsy. We have used the model of syngeneic BMT and simultaneous infection of BALB/c mice with murine CMV for studying the pathogenesis of CMV-IP by controlled longitudinal analysis. A disseminated cytopathic infection of the lungs with fatal outcome was observed only when reconstituting CD8 T cells were depleted. Neither CD8 nor CD4 T cells mediated an immunopathogenesis of acute CMV-IP. By contrast, after efficient hematolymphopoietic reconstitution, viral replication in the lungs was moderate and focal. The histopathological picture was dominated by preferential infiltration of CD8 T cells confining viral replication to inflammatory foci. Notably, after clearance of acute infection, CD62L(lo) and CD62L(hi) subsets of CD44(+) memory CD8 T cells were found to persist in lung tissue. One can thus operationally distinguish an early CMV-positive IP (phase 1) and a late CMV-negative IP (phase 2). According to the definition, phase 2 histopathology would not be diagnosed as a CMV-IP and could instead be misinterpreted as a CMV-induced immunopathology. We document here that phase 1 as well as phase 2 pulmonary CD8 T cells are capable of exerting effector functions and are effectual in protecting against productive infection. We propose that antiviral "stand-by" memory-effector T cells persist in the lungs to prevent virus recurrence from latency.     

Emergence of CD8+ T cells expressing NK cell receptors in influenza A virus-infected mice

Both innate and adaptive immune responses play an important role in the recovery of the host from viral infections. In the present report, a subset of cells coexpressing CD8 and NKR-P1C (NK1.1) was found in the lungs of mice infected with influenza A virus. These cells were detected at low numbers in the lungs of uninfected mice, but represented up to 10% of the total CD8(+) T cell population at day 10 postinfection. Almost all of the CD8(+)NK1.1(+) cells were CD8alphabeta(+)CD3(+)TCRalphabeta(+) and a proportion of these cells also expressed the NK cell-associated Ly49 receptors. Interestingly, up to 30% of these cells were virus-specific T cells as determined by MHC class I tetramer staining and by intracellular staining of IFN-gamma after viral peptide stimulation. Moreover, these cells were distinct from conventional NKT cells as they were also found at increased numbers in influenza-infected CD1(-/-) mice. These results demonstrate that a significant proportion of CD8(+) T cells acquire NK1.1 and other NK cell-associated molecules, and suggests that these receptors may possibly regulate CD8(+) T cell effector functions during viral infection.     

CD94/NKG2A expression is associated with proliferative potential of CD8 T cells during persistent polyoma virus infection

Memory CD8 T cells comprise a critical component of durable immunity because of their capacity to rapidly proliferate and exert effector activity upon Ag rechallenge. During persistent viral infection, memory CD8 T cells repetitively encounter viral Ag and must maintain a delicate balance between limiting viral replication and minimizing immunopathology. In mice infected by polyoma virus, a natural mouse pathogen that establishes long-term persistent infection, the majority of persistence-phase antiviral CD8 T cells express the inhibitory NK cell receptor CD94/NKG2A. In this study, we asked whether CD94/NKG2A expression is associated with Ag-specific recall of polyoma virus-specific CD8 T cells. During the persistent phase of infection, polyoma virus-specific CD8 T cells that express CD94/NKG2A were found to preferentially proliferate; this proliferation was dependent on cognate Ag both in vitro and in vivo. In addition, CD94/NKG2A(+) polyoma-specific CD8 T cells have a markedly enhanced capacity to produce IL-2 upon ex vivo Ag stimulation compared with CD94/NKG2A(-) polyoma-specific CD8 T cells. Importantly, CD94/NKG2A(+) anti-polyoma virus CD8 T cells appear to be essential for Ag-specific recall responses in mice persistently infected by polyoma virus. Because of its higher proliferative potential and capacity to produce IL-2, we propose that the CD94/NKG2A(+) subpopulation represents a less differentiated state than the CD94/NKG2A(-) subpopulation. Identification of proliferation-competent subpopulations of memory CD8 T cells should prove valuable in designing therapeutic vaccination strategies for persistent viral infections.     

CD4(+) T cells are required for the development of cytotoxic CD8(+) T cells during Mycobacterium tuberculosis infection.

The control of acute and chronic Mycobacterium tuberculosis infection is dependent on CD4(+) T cells. In a variety of systems CD8(+) T cell effector responses are dependent on CD4(+) T cell help. The development of CD8(+) T cell-mediated immune responses in the absence of CD4(+) T cells was investigated in a murine model of acute tuberculosis. In vitro and in vivo, priming of mycobacteria-specific CD8(+) T cells was unaffected by the absence of CD4(+) T cells. Infiltration of CD8(+) T cells into infected lungs of CD4(-/-) or wild-type mice was similar. IFN-gamma production by lung CD8(+) T cells in CD4(-/-) and wild-type mice was also comparable, suggesting that emergence of IFN-gamma-producing mycobacteria-specific CD8(+) T cells in the lungs was independent of CD4(+) T cell help. In contrast, cytotoxic activity of CD8(+) T cells from lungs of M. tuberculosis-infected mice was impaired in CD4(-/-) mice. Expression of mRNA for IL-2 and IL-15, cytokines critical for the development of cytotoxic effector cells, was diminished in the lungs of M. tuberculosis-infected CD4(-/-) mice. As tuberculosis is frequently associated with HIV infection and a subsequent loss of CD4(+) T cells, understanding the interaction between CD4(+) and CD8(+) T cell subsets during the immune response to M. tuberculosis is imperative for the design of successful vaccination strategies.     

Immunotherapy with MVA-BN?-HER2 induces HER-2-specific Th1 immunity and alters the intratumoral balance of effector and regulatory T cells

MVA-BN?-HER2 is a new candidate immunotherapy designed for the treatment of HER-2-positive breast cancer. Here, we demonstrate that a single treatment with MVA-BN?-HER2 exerts potent anti-tumor efficacy in a murine model of experimental pulmonary metastasis. This anti-tumor efficacy occurred despite a strong tumor-mediated immunosuppressive environment characterized by a high frequency of regulatory T cells (T(reg)) in the lungs of tumor-bearing mice. Immunogenicity studies showed that treatment with MVA-BN?-HER2 induced strongly Th1-dominated HER-2-specific antibody and T-cell responses. MVA-BN?-HER2-induced anti-tumor activity was characterized by an increased infiltration of lungs with highly activated, HER-2-specific, CD8+CD11c+ T cells accompanied by a decrease in the frequency of T(reg) cells in the lung, resulting in a significantly increased ratio of effector T cells to T(reg) cells. In contrast, administration of HER2 protein formulated in Complete Freund's Adjuvant (CFA) induced a strongly Th2-biased immune response to HER-2. However, this did not lead to significant infiltration of the tumor-bearing lungs by CD8+ T cells or the decrease in the frequency of T(reg) cells nor did it result in anti-tumor efficacy. In vivo depletion of CD8+ cells confirmed that CD8 T cells were required for the anti-tumor activity of MVA-BN?-HER2. Furthermore, depletion of CD4+ or CD25+ cells demonstrated that tumor-induced T(reg) cells promoted tumor growth and that CD4 effector cells also contribute to MVA-BN?-HER2-mediated anti-tumor efficacy. Taken together, our data demonstrate that treatment with MVA-BN?-HER2 controls tumor growth through mechanisms including the induction of Th1-biased HER-2-specific immune responses and the control of tumor-mediated immunosuppression.     

Prognostic value of the immunological phenomena and relationship with clinicopathological characteristics of the tumor--the expression of the early CD69+, CD71+ and the late CD25+, CD26+, HLA/DR+ activation markers on T CD4+ and CD8+ lymphocytes in squamous cell laryngeal carcinoma. Part II

One of the most important challenges in contemporary oncology is to find objective biomarkers of tumor aggressiveness, which help to identify more invasive phenotypes of the carcinoma. The purpose of this study was to investigate the relationships between the early and the late activation markers expression on T CD4(+) and CD8(+) cells subpopulations and certain clinicopathological characteristics of the neoplastic infiltration in order to determine their role as biomarkers for tumor behavior in squamous cell laryngeal carcinoma. Analysis of the early (CD69(+), CD71(+)) and the late activation antigens (CD25(+) (high), CD26(+), HLA/DR(+)) expression on T CD4+ and CD8(+) lymphocytes by cytofluorymetry in 55 patients treated for squamous cell laryngeal carcinoma was performed. Clinicomorphological analysis on the basis of TNM criteria and tumor front grading, which included tumor-related features and adjacent stroma-related characteristics of the peripheral edge of infiltration was carried out. The relationships between the activation markers expression and parameters of tumor aggressiveness were investigated. Our work revealed statistically significant differences in the expression of the studied activation markers on T cells with regard to certain clinicomorphological features. The expressions of CD69(+) and CD71(+) antigens on T CD3(+)CD4(+) and CD3(+)CD8(+) cells as well as CD4(+)HLA/DR(+) markers were higher for pT3 and pT4 tumors, in comparison with pT2 carcinomas. Moreover, tumors with the smallest number of TFG points were characterized by significantly lower values of the average expression of CD3(+)CD69(+) and CD3(+)CD71(+) as well as CD4(+)HLA/DR(+) markers on T lymphocytes. In addition, more aggressive and deeply infiltrating laryngeal carcinomas were most often characterized by significantly higher values of the average expression of CD69(+) and CD71(+) antigens on CD8(+) as well as HLA/DR(+) markers on CD4(+). Our study confirmed the implication of the early and the late activation antigens expression on CD4(+) and CD8(+) T lymphocytes in clinicomorphological parameters of the tumor, especially TFG total score and depth of invasion, and their importance as indicators of the invasive phenotype of laryngeal carcinoma.