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Immunophenotypic analysis of the tCR-Vβ repertoire in 98 persistent expansions of cd3⁺/tCR-αβ⁺ large granular lymphocytes -: Utility in assessing clonality and insights into the pathogenesis of the disease

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AMERICAN JOURNAL OF PAtHOLOGY 2001年第5期159卷 1861-1868页

作者： Lima, M Almeida, J Santos, AH teixeira, MDA Alguero, MDC Queirós, ML Balanzategui, A Justiça, B Gonzalez, M Miguel, JFS Orfao, A Hosp Geral Santo Antonio Serv Hematol Clin Unidade Citometria P-4050 Oporto Portugal Hosp Univ Salamanca Serv Citometria Salamanca Spain Hosp Univ Salamanca Serv Hematol Salamanca Spain Univ Salamanca Ctr Invest Canc E-37008 Salamanca Spain

At present, a major challenge in the initial diagnosis of leukemia of large granular lymphocytes (LGLs) is to establish the clonal nature of the expanded population. in the present study we have analyzed by flow cytometry immunophenotyping the tCR-V beta repertoire of 98 consecutive cases of persistent expansions of cd4(+) or cd8(+bright)cd3(+)/tCR-alpha beta (+) LGLs and compared the results with those obtained in molecular studies of tCR-beta gene rearrangements. Fifty-eight cases were considered to be monoclonal in molecular studies whereas in the remaining 40 cases there was no evidence for monoclonality (11 cases were considered oligoclonal and 29 polyclonal). the tCR-V beta repertoire was biased to the preferential use of one or more tCR-V beta families in 96% of cases, a total of 124 tCR-V beta expansions being diagnosed: one tCR-V beta expansion in 71 cases and two or more tCR-V beta expansions in 23 cases. the highest tCR-V beta expansion observed in each case was higher among monoclonal (74 +/- 19%) as compared to nonmonoclonal cases (24 +/- 14%) (P = 0.001), as did the fraction of LGLs that exhibited a tCR-V beta -restricted pattern (86 +/- 16% and 42 +/- 23%, respectively;P = 0.0001);by contrast, the proportion of cases displaying more than one tCR-V beta expansion was higher in the latter group: 7% versus 48%, respectively (P = 0.001). Results obtained in oligoclonal cases were intermediate between those obtained in polyclonal and monoclonal cases and similar results were observed for cd4(+) as for cd8(+bright) t-cell expansions. tCR-V beta families expressed in cd8(+bright) t-cell-LGL proliferations showed a pattern of distribution that mimics the frequency at which the individual tCR-V beta families are represented in normal peripheral blood t cells. Assuming that a given proliferation of LGLs is monoclonal whenever there is an expansion of a given tCR-V beta family of at least 40% of the total cd4(+) or cd8(+bright) t-cell. compartment, we were able

关键词： cd4阳性t淋巴细胞/生理学 cd8阳性t淋巴细胞/生理学克隆细胞免疫表型分型白血病实验性/病因学白血病实验性/免疫学白血病实验性/病理学受体抗原 t细胞 α-β/遗传学参考值 t淋巴细胞/病理学 t淋巴细胞/生理学青少年成年人老年人老年人 80以上儿童女(雌)性人类男(雄)性中年人

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Long-term survival but impaired homeostatic proliferation of naive t cells in the absence of p56^lck

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SCIENCE 2000年第5489期290卷 127-131页

作者： Seddon, B Legname, G tomlinson, P Zamoyska, R Natl Inst Med Res Div Mol Immunol London NW7 1AA England Univ Calif San Francisco Inst Neurodegenerat Dis San Francisco CA 94143 USA

Interactions between the t cell receptor (tCR) and major histocompatibility complex antigens are essential for the survival and homeostasis of peripheral t lymphocytes, However, little is known about the tCR signaling events that result from these interactions. the peripheral t cell pool of p56(lck) (lck)-deficient mice was reconstituted by the expression of an inducible lck transgene. Continued survival of peripheral naive t cells was observed for long periods after switching off the transgene, Adoptive transfer of t cells from these mice into t lymphopoienic hosts confirmed that t cell survival was independent of lck but revealed its essential role in tCR-driven homeostatic proliferation of naive t cells in response to the t cell-deficient host environment. these data suggest that survival and homeostatic expansion depend on different signals.

关键词： cd4阳性t淋巴细胞/免疫学 cd4阳性t淋巴细胞/生理学 cd8阳性t淋巴细胞/免疫学 cd8阳性t淋巴细胞/生理学细胞分裂细胞存活多西环素/药理学基因表达内环境稳定淋巴细胞活化淋巴细胞计数淋巴细胞特异蛋白质酪氨酸激酶p56(Lck)/遗传学淋巴细胞特异蛋白质酪氨酸激酶p56(Lck)/生理学淋巴细胞输注淋巴组织/细胞学淋巴组织/免疫学小鼠基因敲除小鼠转基因磷酰化原癌基因蛋白质c-bcl-2/遗传学原癌基因蛋白质c-bcl-2/代谢受体抗原 t细胞/代谢信号传导 t淋巴细胞亚群/免疫学 t淋巴细胞亚群/生理学 t淋巴细胞/免疫学 t淋巴细胞/生理学 t淋巴细胞/移植胸腺/细胞学胸腺/免疫学转基因动物小鼠

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cd4 t lymphocytes are primed to express Fas ligand by cd4 cross-linking and to contribute to cd8 t-cell apoptosis via Fas/FasL death signaling pathway

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BLOOD 2000年第1期96卷 195-202页

作者： tateyama, M Oyaizu, N McCloskey, tW than, S Pahwa, S NYU N Shore Univ Hosp Dept Pediat Sch MedDiv Allergy Immunol Manhasset NY 11030 USA Tokyo Med & Dent Univ Dept Retroviral Regulat Div Med Res Bunkyo Ku Tokyo Japan

cd4 molecules serve as coreceptors for the t-cell receptor (tCR)/cd3 complex that are engaged coordinately with tCR and facilitate antigen-specific t-cell activation leading to interleukin 2 (IL-2) production and proliferation. However, cross-ligation of cd4 molecules prior to tCR stimulation has been shown to prime cd4 t cells to undergo apoptosis. Although in vivo and in vitro experiments have implicated the involvement of Fas/FasL interaction in this cd4 cross-linking (cd4XL)-induced apoptosis, detailed mechanisms to account for cell death induction have not been elucidated. In the present study, we demonstrate that cd4XL in purified t cells not only led to Fas up-regulation but also primed cd4 t cells to express Fast upon cd3 stimulation and rendered the t cells susceptible to Fas mediated apoptosis. Notably, in addition to cd4(+) t cells, cd4XL-induced sensitization for apoptosis was observed in cd8(+) t cells as well and was associated with Bcl-x down-modulation. Both cd4 and cd8 t-cell subsets underwent apoptosis following cell-cell contact with FasL(+) cd4 t cells. cd28 costimulation abrogated cd4XL/ cd3-induced apoptosis with restoration of IL-2 production and prevented Bcl-x down-modulation. As cd4 molecules are the primary receptors for human immunodeficiency virus 1 (HIV-1), we conclude that HIV-1 envelope mediated cd4XL can lead to the generation of FasL-expressing cd4+ t cells that can lead to apoptosis of cd4 as well as cd8 t cells. these findings implicate a novel mechanism for cd8 t-cell depletion in HIV disease. (Blood. 2000;98:195-202) (C) 2000 by the American Society of Hematology.

关键词：抗原 cd/免疫学抗原 cd3/免疫学抗原 cd4/免疫学抗原 cd95/免疫学细胞凋亡/免疫学 cd4阳性t淋巴细胞/免疫学 cd4阳性t淋巴细胞/生理学 cd8阳性t淋巴细胞/细胞学 cd8阳性t淋巴细胞/免疫学 cd8阳性t淋巴细胞/生理学 Fas配体蛋白质干扰素Ⅱ型/生物合成淋巴细胞活化膜糖蛋白类/遗传学膜糖蛋白类/免疫学 RNA 信使/遗传学逆转录聚合酶链反应信号传导/免疫学肿瘤坏死因子α/生物合成人类

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Immune-recovery uveitis in patients with cytomegalovirus retinitis taking highly active antiretroviral therapy

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AMERICAN JOURNAL OF OPHtHALMOLOGY 2000年第1期130卷 49-56页

作者： Robinson, MR Reed, G Csaky, KG Polis, MA Whitcup, SM NEI NIH Bethesda MD 20892 USA NIAID NIH Bethesda MD 20892 USA

PURPOSE: to investigate the clinical features associated with immune recovery in human immunodeficiency virus (HIV)-infected patients with cytomegalovirus retinitis who are taking highly active antiretroviral therapy. MEtHODS: Sixteen patients were evaluated prospectively at the National Eye Institute, Bethesda, Maryland. Evaluation included a medical history and a complete ophthalmologic examination. the examination included best corrected visual acuity score measured by means of logarithmic charts, slit-lamp biomicroscopy, dilated retinal examination, retinal photography, and fluorescein angiography. Immune-recovery uveitis was defined as the ocular inflammation associated with clinical immune the presence of moderate to severe macular edema on fluorescein angiography and vitreous haze (P less than or equal to .001). CONCLUSIONS: Immune-recovery uveitis is an important cause of visual morbidity in HIV-infected patients with cytomegalovirus retinitis in the era of highly active antiretroviral therapy. Although immune recovery associated with highly active antiretroviral therapy has allowed some patients to discontinue specific anticytomegalovirus therapy, the rejuvenated immune response can be associated with sight-threatening inflammation. Published by Elsevier Science Inc.

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Highly active antiretroviral therapy-related immune recovery in AIDS patients with cytomegalovirus retinitis

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OPHtHALMOLOGY 2000年第5期107卷 877-881页

作者： Macdonald, JC Karavellas, MP torriani, FJ Morse, LS Smith, IL Reed, JB Freeman, WR Univ Texas Hlth Sci Ctr Dept Ophthalmol San Antonio TX 78284 USA Univ Calif San Diego Shiley Eye Ctr Dept Ophthalmol La Jolla CA 92093 USA Univ Calif San Diego Dept Med La Jolla CA 92093 USA Univ Calif Davis Dept Ophthalmol Davis CA 95616 USA Univ Calif San Diego Dept Pediat La Jolla CA 92093 USA

Objective: to characterize cytomegalovirus (CMV) retinitis in human immunodeficiency virus (HIV)-infected patients who demonstrate immune recovery while receiving highly active antiretroviral therapy (HAARt), Design: Consecutive, noncomparative case series. Participants: twenty-two HIV-positive patients, from two institutions, with a history of CMV retinitis, and with elevated cd4 cell counts after HAARt. Main Outcome Measures: Duration of healed CMV retinitis without anti-CMV therapy, cd4 cell count, and HIV viral load. Intervention: Discontinuation of anti-CMV therapy after persistent elevation of cd4 cell count over 50 cell/mm(3) (median, 161/mm(3);range, 85-408/mm(3)). Results: the median period of healed CMV retinitis without anti-CMV therapy was 72 weeks (range, 33-116 weeks). Nineteen of 22 patients were still healed without anti-CMV therapy at study end. the three patients with CMV retinitis progression simultaneously had HAARt, fail with cd4 cell counts of 37, 35, and 47/mm(3). Conclusions: HIV-positive patients with CMV retinitis, who demonstrate a sustained HAARt-induced elevation of cd4 cell count on two consecutive counts 3 months apart and whose retinitis remains healed on anti-CMV therapy for greater than 4 months, are likely to remain healed if the anti-CMV therapy is withdrawn, It is important to monitor these patients with indirect ophthalmoscopy because HAARt failure may occur and allow CMV retinitis reactivation. (C) 2000 by the American Academy of Ophthalmology.

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Role of cysteinyl leukotrienes in cd⁴⁺ t cell-driven late allergic airway responses

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JOURNAL OF PHARMACOLOGY AND EXPERIMENtAL tHERAPEUtICS 2000年第2期293卷 410-416页

作者： Hojo, M Suzuki, M Maghni, K Hamid, Q Powell, WS Martin, JG McGill Univ Royal Victoria Hosp Meakins Christie Labs Montreal PQ H3A 1A1 Canada

Cysteinyl leukotrienes (cys Lts) play an important role in late responses to allergen challenge of actively sensitized rats. the aim of this study was to determine whether t cell-dependent late airway responses (LARs) also are mediated by cys Lts. to do this we tested the effects of the selective and potent LtD4 antagonist pranlukast on airway responses to ovalbumin (OVA) challenge of naive recipients of cd4(+) t cells isolated from the cervical lymph nodes of OVA-sensitized donor rats. cd4(+) t cells (5 million) were purified by immunomagnetic separation and administered i.p. 2 days before OVA challenge. the pulmonary resistance was measured for 8 h after challenge and bronchoalveolar lavage (BAL) was performed for analysis of leukocytes and major basic protein-positive cells. the LAR, determined as the area under the curve of pulmonary resistance against time from 3 to 8 h after challenge, was 8.9 +/- 1.79 cm H2O/ml/s x min after OVA compared with 2.8 +/- 0.50 cm H2O/ml/s x min (P<.01) after OVA and pranlukast treatment. the total cell count in BAL was not significantly greater in the OVA challenged group (3.55 +/- 0.41 x 10(6) cells) compared with the OVA- and pranlukast-treated group (2.65 +/- 0.45 x 10(6) cells). However, lymphocytes and eosinophils were reduced in numbers by pranlukast. Interleukin-5 mRNA-positive cells were diminished by 50% in pranlukast-treated animals. In conclusion, pranlukast inhibits LAR, BAL eosinophilia, and Interleukin-5 expression in rats with adoptively transferred LAR, indicating an important role for cys Lts in these t cell-driven responses.

关键词：过继转移气道阻力/药物作用变应原/药理学抗变态反应药/药理学血蛋白质类/代谢支气管肺泡灌洗液/细胞学 cd4阳性t淋巴细胞/生理学色酮类/药理学嗜酸细胞颗粒蛋白质类嗜酸粒细胞增多/病理学原位杂交白细胞介素5/代谢白三烯拮抗剂/药理学白三烯类/生理学淋巴结/细胞学淋巴结/药物作用大鼠近交BN 呼吸超敏反应/代谢呼吸超敏反应/病理学呼吸超敏反应/病理生理学核糖核酸酶类动物男(雄)性大鼠

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Activation of p38 mitogen-activated protein kinase in vivo selectively induces apoptosis of cd8⁺ but not cd4⁺ t cells

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MOLECULAR AND CELLULAR BIOLOGY 2000年第3期20卷 936-946页

作者： Merritt, C Enslen, H Diehl, N Conze, D Davis, RJ Rincón, M Univ Vermont Dept Med Immunobiol Program Burlington VT 05405 USA Univ Massachusetts Sch Med Dept Biochem & Mol Biol Program Mol Med Worcester MA 01605 USA Howard Hughes Med Inst Worcester MA 01605 USA

cd4(+) and cd8(+) t cells play specific roles during an immune response. Different molecular mechanisms could regulate the proliferation, death, and effector functions of these two subsets of t cells. the p38 mitogen-activated protein (MAP) kinase pathway is induced by cytokines and environmental stress and has been associated with cell death and cytokine expression. Here me report that activation of the p38 MAP kinase pathway in vivo causes a selective loss of cd8(+) t cells due to the induction of apoptosis. In contrast, activation of p38 MAP kinase does not induce cd4(+) t-cell death. the apoptosis of cd8(+) t cells is associated with decreased expression of the antiapoptotic protein Bcl-2. Regulation of the p38 MAP kinase pathway in t cells is therefore essential for the maintenance of cd4/cd8 homeostasis in the peripheral immune system. Unlike cell death, gamma interferon production is regulated by the p38 MAP kinase pathway in bath cd4(+) and cd8(+) t cells, thus, specific aspects of cd4(+) and cd8(+) t-cell function are differentially controlled by the p38 MAP kinase signaling pathway.

关键词：细胞凋亡/生理学 cd4阳性t淋巴细胞/细胞学 cd4阳性t淋巴细胞/免疫学 cd4阳性t淋巴细胞/生理学 cd8阳性t淋巴细胞/细胞学 cd8阳性t淋巴细胞/免疫学 cd8阳性t淋巴细胞/生理学钙-钙调素依赖性蛋白激酶类细胞培养的酶激活流式细胞术基因表达调控淋巴结/免疫学淋巴细胞活化 MAP激酶激酶6 小鼠转基因丝裂原激活蛋白激酶激酶类/遗传学丝裂原激活蛋白激酶激酶类/代谢丝裂原激活蛋白激酶类/代谢原癌基因蛋白质c-bcl-2/遗传学逆转录聚合酶链反应脾/免疫学 p38丝裂原活化蛋白激酶类动物小鼠

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Altered lymphoid development in mice deficient for the mAF4 proto-oncogene

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BLOOD 2000年第2期96卷 705-710页

作者： Isnard, P Coré, N Naquet, P Djabali, M CNRS INSERM Ctr Immunol F-13288 Marseille 9 France

Some chromosomal translocations in acute leukemias involve the fusion of the trithorax-related protein MII (also called HRX, All1, Htrx,) with a variety of heterologous proteins. In acute lymphoblastic leukemia associated with the t(4;11)(q21;q23) translocation, the 4q21 gene that fuses with MII is AF4, to gain insight into the potential role of AF4 in leukemogenesis and development, this gene was inactivated by homologous recombination in mice. As expected from the tissue distribution of the AF4 transcript, development of both B and t cells is affected in AF4 mutant mice. A severe reduction of the thymic double positive cd4/cd8 (cd4(+)/cd8(+)) population was observed;in addition most double- and single-positive cells expressed lower levels of cd4 and cd8 coreceptors, Most importantly, the reconstitution of the double-positive compartment by expansion of the double-negative cell compartment was severely impaired in these mutant mice. In the bone marrow pre-B and mature B-cell numbers are reduced. these results demonstrate that the function of the mAF4 gene is critical for normal lymphocyte development. this raises the possibility that the disruption of the normal AF4 gene or its association with MII function by translocation may orient the oncogenic process toward the lymphoid lineage. this represents the first functional study using a knock-out strategy on one of the Mil partner genes in translocation-associated leukemias.

关键词： B淋巴细胞/生理学骨髓细胞/生理学 cd4阳性t淋巴细胞/生理学 cd8阳性t淋巴细胞/生理学 DNA结合蛋白质类/缺乏 DNA结合蛋白质类/遗传学糖皮质激素类/药理学血细胞生成淋巴细胞计数淋巴细胞/生理学小鼠近交BALBC 小鼠基因敲除诱变髓淋巴细胞性白血病蛋白质核蛋白质类/缺乏核蛋白质类/遗传学原癌基因 t淋巴细胞/生理学转录因子转染易位遗传动物女(雌)性男(雄)性小鼠

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Long-term posterior and anterior segment complications of immune recovery uveitis associated with cytomegalovirus retinitis

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AMERICAN JOURNAL OF OPHtHALMOLOGY 2000年第1期130卷 57-64页

作者： Karavellas, MP Song, M Macdonald, JC Freeman, WR Univ Calif San Diego Dept Ophthalmol Shiley Eye Ctr La Jolla CA 92093 USA

PURPOSE: to identify and describe long-term posterior and anterior segment complications of immune recovery uveitis in patients with inactive cytomegalovirus retinitis who are undergoing highly active antiretroviral therapy-mediated recovery of immune function. MEtHODS: A prospective cohort study at a university medical center. twenty-nine eyes of 21 patients with immune recovery uveitis and inactive cytomegalovirus retinitis were followed for 14.5 to 116 weeks (median, 43 weeks) after diagnosis of immune recovery uveitis, RESULtS: Nine eyes of nine patients developed visually important complications involving the posterior segment, anterior segment, or a combination of both. Posterior segment complications included severe proliferative vitreoretinopathy in three eyes and spontaneous vitreous hemorrhage from avulsion of a blood vessel secondary to tory syndrome that may result in complications months to years after the onset of inflammation. (C) 2000 by Elsevier Science Inc. All rights reserved.

关键词：眼前半段/病理学抗逆转录病毒治疗高效抗病毒药/治疗应用 cd4淋巴细胞计数 cd4阳性t淋巴细胞/生理学白内障/病因学白内障/病理学队列研究巨细胞病毒视网膜炎/并发症巨细胞病毒视网膜炎/药物疗法巨细胞病毒视网膜炎/免疫学巨细胞病毒视网膜炎/病毒学免疫系统/生理学虹膜炎/病因学虹膜炎/病理学虹膜炎/外科学前瞻性研究复发时间因素葡萄膜炎/并发症葡萄膜炎/药物疗法葡萄膜炎/免疫学葡萄膜炎/病毒学视敏度玻璃体视网膜病增生性/病因学玻璃体视网膜病增生性/病理学玻璃体视网膜病增生性/外科学玻璃体出血/病因学玻璃体出血/病理学玻璃体出血/外科学成年人人类男(雄)性

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Expression of 1,25-dihydroxyvitamin D₃ receptor in the immune system

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ARCHIVES OF BIOCHEMIStRY AND BIOPHYSICS 2000年第2期374卷 334-338页

作者： Veldman, CM Cantorna, Mt DeLuca, HF Univ Wisconsin Coll Agr & Life Sci Dept Biochem Madison WI 53706 USA

In addition to its role in calcium and skeletal homeostasis, there is increasing evidence that the hormonal form of vitamin D-,D- 1,25-dihydroxyvitamin D-3, appears to serve as a modulator of the immune system. We have determined the level of the 1,25-dihydroxyvitamin D-3 receptor (VDR) in resting and activated lymphocytes by immuno- and ligand-binding assays. As expected from previous work, the total t lymphocyte population contains VDR whose levels are increased when activated and treated with 1,25-dihydroxyvitamin D-3. Surprisingly, the highest concentrations of VDR are found in cd8 lymphocytes, although significant amounts are also present in cd4 lymphocytes. Furthermore, B lymphocytes do not contain detectable amounts of VDR. Cells of the monocyte/macrophage lineage possess small amounts of VDR that are not affected by activation but are increased by treatment with 1,25-dihydroxyvitamin D-3. these results suggest that cd8 lymphocytes may be a major site of 1,25-dihydroxyitamin D-3 action, while B lymphocytes are likely not directly regulated by 1,25-dihydroxyvitamin D-3. (C) Academic Press.

关键词： B淋巴细胞/细胞学 B淋巴细胞/药物作用 B淋巴细胞/生理学 cd4阳性t淋巴细胞/生理学 cd8阳性t淋巴细胞/生理学骨化三醇/药理学细胞分裂细胞培养的动力学淋巴细胞活化小鼠近交系受体骨化三醇/生物合成 t淋巴细胞/细胞学 t淋巴细胞/药物作用 t淋巴细胞/生理学动物人类小鼠

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