淋巴管阻断对大鼠睾丸生精上皮的影响

用外科手术的方法将正常成年ＳＤ大鼠双侧睾丸淋巴管阻断后，观察术后不同时间睾丸生精上皮的组织学变化。术后２天出现生精上皮的形态改变，生精细胞排列松散，部分精子细胞脱落，聚集在管腔部位。术后６天多数曲细精管的精子细胞全部脱落。假手术对照组，睾丸生精上皮具有正常的形态结构。     

大鼠睾丸间质细胞发育的形态学研究及立体学定量分析

对６３例胚胎期至生后不同发育时期Ｗｉｓｔａｒ大鼠睾Ｌｅｙｄｉｇ细胞进行光镜、电镜观察及立体学分析。结果表明：１．大鼠睾丸Ｌｅｙｄｉｇ细胞从胚胎１５ｄ出现，１７ｄ初具分泌类固醇激素的结构特点。２．Ｌｅｙｄｉｇ细胞存在两个细胞增殖峰，一个在胚胎１７￣１９ｄ，另一个在生后４周。两峰之间有一个低谷（约在生后第２周）。从胚胎至生后，细胞核的体密度逐渐减小，细胞质、线粒体及滑面内质网的体密度逐渐增大，脂滴的体     

小白鼠睾丸发育分化的组织学及组织化学观察

本文报道用组织学和组织化学方法观察小鼠睾丸胚胎天时睾丸即已有SDH活性,生后以间质细胞和精母细胞反应最强。4.AlP反应在胚胎时以生殖母细胞较强,生后以界膜反应较强。5.支持细胞、间质细胞AcP活性较强。6.生殖细胞和支持细胞5′-Nase呈阳性,间质反应极微。7.胚胎时,睾丸各种细胞ATPase反应均为阴性。生后界膜、支持细胞和生殖细胞渐出现活性且不断增强。8.胚胎14天睾丸内NSE即有微弱的反应,生后主要以间质细胞反应强烈。至生后的发育。结果表明:1.核糖核酸以代谢旺盛的生殖母细胞最为丰富,支持细胞也较多,精子细胞较少。2.胚胎14天时睾丸各种细胞均有丰富的糖原颗粒,16天起仅分布于间质,性索内糖原阴性。3.胚胎14     

白细胞介素4基因转移的肿瘤细胞体内增强巨噬细胞功能的实…

观察了小鼠接种高分泌ＩＬ－４的黑色素瘤细胞后腹腔内巨噬细胞功能的变化，荷瘤小鼠腹腔内巨噬细胞杀伤性在荷瘤后第４天及第１２天出现两个高峰，荷瘤后第１５天小鼠腹腔内巨噬细胞的吞噬能力及ＭＨＣⅡ类抗原的表达增强，抗原提呈能力增高，荷瘤后第４天及第１２天腹腔内巨噬细胞经诱导的ＩＬ－１水平出现两个高峰，经诱导的ＴＮＦ水平变化不明显，此实验结果表明，高分泌ＩＬ－４的黑色素瘤细胞体内生长受抑制的原因之一是ＩＬ－     

白细胞介素4基因转移的肿瘤细胞体内增强巨噬细胞功能的实验研究

观察了小鼠接种高分泌ＩＬ－４的黑色素瘤细胞后腹腔内巨噬细胞功能的变化。荷瘤小鼠腹腔内巨噬细胞杀伤性在荷瘤后第４天及第１２天出现两个高峰；荷瘤后第１５天小鼠腹腔内巨噬细胞的吞噬能力及ＭＨＣⅡ类抗原的表达增强，抗原提呈能力增高；荷瘤后第４天及第１２天腹腔内巨噬细胞经诱导的ＩＬ－１水平出现两个高峰，经诱导的ＴＮＦ水平变化不明显。此实验结果表明，高分泌ＩＬ－４的黑色素瘤细胞体内生长受抑制的原因之一是ＩＬ－４基因的导入及ＩＬ－４的分泌使体内巨噬细胞的抗原提呈能力增强及杀伤活性的显著提高，因而使机体抗肿瘤免疫功能得以增强。     

42 大鼠出生前后睾丸中两种中间丝蛋白的免疫组化观察

<正>本文旨在探讨细胞角蛋白和波蛋白在大鼠发育中睾丸内的表达,特别是在支持细胞内的表达.阴栓龄17～20天的Wistar胎鼠和生后1、5、10、15、20和40天的幼鼠购自中国医学科学院实验动物中心,生后1天的仔鼠及胎鼠取右侧睾丸于4%多聚甲醛中浸泡固定.生后5天以上的幼鼠先经心脏灌流固定,再取睾丸浸泡固定.石蜡包埋后组织块作5μm厚切片.PAP法或ABC法显示细胞角蛋白,Biotin-Streptoavidin法显示波蛋白.兔抗牛角蛋白多克隆抗体和小鼠抗猪波蛋白单克隆抗体均为DAKO公司产品.结果如下:(1)17天龄胎鼠睾丸可见两种中间纵蛋白的表达.角蛋白主要见于表面上皮,纵隔及其附近的睾丸索(testicular cords)内的上皮细胞以及其它睾丸索的支持细胞.在纵隔以外的睾丸索,角蛋白的反应弱,只见于支持细胞的核下区.波蛋白分布于白膜、问质及血管内皮.在睾丸索内则分布在支持细胞的核下区.(2)随年龄增长,睾丸索内支持细胞的角蛋白表达减弱.纵隔及其附近的睾丸索除外,它们发展成为直细精管和睾丸网.生后15天以后,睾丸索(曲细精管)支持细胞的角蛋白用PAP法已不能显示.(3)用ABC法,在出生后15天以上的曲细精管内确可见到少数角蛋白阳性细胞,大多数支持细胞的波蛋白表达增强.本结果提示,支持细胞可能有两种类型.随着发育,大多数支持细?     

SARS男性患者睾丸组织的病理改变

目的研究严重急性呼吸综合征(SARS)男性患者睾丸组织的病理变化。方法对5例SARS死亡患者的睾丸组织进行常规组织学、TUNEL及免疫组化染色。结果5例SARS患者睾丸生精小管基膜增厚、纤维化,生精上皮坏死、脱落,睾丸中几乎未见精子。生精细胞凋亡大量增加(P<0.05),间质充血,睾丸组织中有明显的白细胞浸润。CD3 T淋巴细胞、CD68 巨噬细胞较对照组明显增多(P<0.05),并浸润到生精小管中。结论SARS导致男性患者并发睾丸炎。     

生精细胞凋亡的FCM、TUNEL检测及相关基因表达

目的及方法:生精细胞凋亡对于保证精子的正常数目具重要意义,已表明生精细胞凋亡与年龄、生精上皮时相及调控基因有关。为明确生精细胞凋亡的规律及相关基因的表达,应用免疫组化、TUNEL染色(缺口末端标记法)、FCM(流式细胞术)及透射电镜(TEM)等技术对生后4天、8天、12天、16天、20天、24天、28天、32天、60天、3个月及5个月的SD大鼠的生精细胞进行了研究。将动物的每侧睾丸分别制备石蜡、超薄切片及单细胞悬液以备观察。结果及结论:1凋亡率(FCM):在生后的4天组到32天组,生精细胞的凋亡率逐步升高,32天达高峰之后,随着性成熟而下降。2…     

白细胞介素4基因转移的肿瘤细胞体内诱导杀伤细胞活性及细胞因子产生的实验研究

观察了小鼠接种高分泌ＩＬ－４的黑色素瘤细胞后体内免疫效应细胞（包括ＣＴＬ、ＮＫ细胞及ＬＡＫ细胞）抗肿瘤活性及其分泌的细胞因子（包括ＩＬ－２、ＩＦＮ－ｒ、ＴＮＦ及ＧＭ－ＣＳＦ）水平的变化，荷瘤后第１５天小鼠脾细胞ＮＫ活性及经诱导后的ＬＡＫ活性有所降低，而经诱导后的ＣＴＬ杀伤活性显著升高，荷瘤小鼠腹腔内巨噬细胞杀伤活性及其分泌的ＩＬ－１水平也明显升高；在荷瘤后第４天及第１２天出现两个高峰，实验结果表明，高分泌ＩＬ－４的黑色素瘤细胞体内生长受抑制是因为ＩＬ－４基因的导入及ＩＬ－４的分泌使体内ＣＴＬ及巨噬细胞杀伤活性提高，因而使机体抗肿瘤免疫功能得以增强。     

兔视网膜中P物质样免疫反应神经元的发育

本实验用免疫细胞化学ABC法,研究了成年、新生和生后兔视网膜中P物质(SP)样免疫反应神经元的定位和发育。结果表明,成年兔视网膜SP样免疫反应细胞胞体位于内核层和节细胞层,胞突分布在内网层的第1、3、5亚层,偶见于视神经纤维层。细胞密度以视纹最高,从视纹向背腹视网膜边缘区密度渐变小。在新生兔视网膜已有SP阳性胞体和胞突出现,胞体主要位于节细胞层,突起在内网层第5亚层,但未形成连续网层,在第1亚层很少,第3亚层未见SP阳性突起。SP阳性细胞密度从新生到生后第4天增加,生后第6天到第12天细胞密度渐下降。生后第12天SP阳性胞体主要位于内核层。生后第20天,SP阳性细胞的形态、密度与分布已接近成年水平。上述结果提示,在兔视网膜中SP样免疫反应胞体和突起在生前已出现,生后继续发育,到生后20天后其形态发育已接近成熟。     

Functional and phenotypic characteristics of testicular macrophages in experimental autoimmune orchitis

Testicular inflammation with compromised fertility can occur despite the fact that the testis is considered an immunoprivileged organ. Testicular macrophages have been described as cells with an immunosuppressor profile, thus contributing to the immunoprivilege of the testis. Experimental autoimmune orchitis (EAO) is a model of organ-specific autoimmunity and testicular inflammation. EAO is characterized by an interstitial inflammatory mononuclear cell infiltration, damage of the seminiferous tubules and germ cell apoptosis. Here we studied the phenotype and functions of testicular macrophages during the development of EAO. By stereological analysis, we detected an increased number of resident (ED2+) and non-resident (ED1+) macrophages in the testicular interstitium of rats with orchitis. We showed that this increase was mainly due to monocyte recruitment. The in vivo administration of liposomes containing clodronate in rats undergoing EAO led to a reduction in the number of testicular macrophages, which correlated with a decreased incidence and severity of the testicular damage and suggests a pathogenic role of macrophages in EAO. By immunohistochemistry and flow cytometry we detected an increased number of testicular macrophages expressing MHC class II, CD80 and CD86 costimulatory molecules in rats with orchitis. Also, testicular macrophages from rats with EAO showed a higher production of IFNgamma (ELISA). We conclude that testicular macrophages participate in EAO development, and the ED1+ macrophage subset is the main pathogenic subpopulation. They stimulate the immune response through the production of pro-inflammatory cytokines and antigen presentation and thus activation of T cells in the target organ.     

Macrophages activated by Listeria monocytogenes induce organ-specific autoimmunity.

We have previously reported an experimental autoimmune model induced by the local infection of Listeria monocytogenes. The unilateral inoculation of virulent Listeria into a testis of a normal mouse induced a delayed-type hypersensitivity response against testicular antigen and caused autoimmune orchitis in the contralateral testis. The orchitis was transferred to naive mice by T cells from the intratesticularly infected mice. In this paper, we demonstrated that avirulent Listeria, which lacks the expression of listeriolysin O, failed to induce any anti-testicular responses or contralateral orchitis even when it was inoculated at a high dose into the testis. Furthermore, the intraperitoneal inoculation of virulent Listeria with testicular antigen induced the anti-testicular responses and orchitis although intraperitoneal inoculation of testicular antigen with avirulent Listeria failed to induce them. The difference between virulent and avirulent Listeria in the induction of anti-testicular responses was supposed to be dependent on the difference in macrophage activation by the two bacterial strains because, first, the anti-testicular responses were elicited in normal mice when macrophages from virulent Listeria-infected mice were intraperitoneally transferred with testicular antigen although no viable bacteria were detected from the macrophages, and secondly, in contrast, the intraperitoneal co-inoculation of macrophages from avirulent Listeria-infected mice and testicular antigen failed to elicit any anti-testicular responses. Finally, we found that the virulent Listeria-induced macrophages expressed a higher level of CD80 (B7-1) and CD86 (B7-2) molecules than did the avirulent Listeria-induced macrophages and naive peritoneal macrophages. These results thus suggest that virulent Listeria activates macrophages to induce autoreactive T cells while avirulent Listeria does not. The up-regulation of B7 molecules by virulent Listeria infection is a candidate of the mechanism for the activation of autoreactive T cells.     

Effect of testicular macrophage conditioned media from rats with autoimmune orchitis on Leydig cell function

PROBLEM: The aim of this study was to investigate the influence of immune-activated testicular macrophages obtained from rats with autoimmune orchitis (EAO) on Leydig cell steroidogenesis. METHOD OF STUDY: EAO was induced in rats by active immunization with testis homogenate and adjuvants. Testicular and peritoneal macrophages from rats with EAO were isolated and cultured for 24 hr. Testosterone (T) production by purified Leydig cells incubated in vitro with macrophage-conditioned media (CM) from rats with EAO or control rats was measured. RESULTS: An increase in T production by Leydig cells incubated with CM from testicular, but not peritoneal, macrophages of rats with EAO was observed. This increase was dose-dependent up to a concentration of 30% CM; proportions higher than 35% exhibited an inhibitory effect. CONCLUSIONS: Immune-activated testicular macrophages obtained from rats with EAO induced both stimulatory and inhibiting steroidogenic effects on Leydig cells in vitro and not the exclusively inhibitory action that has widely been attributed to activated macrophages. This dual effect probably depends on the ability of these cells to synthesize different molecules that may exert opposite effects.     

Quantification of immunocompetent cells in testicular germ cell tumours

The immunocompetent cells present in the different histological patterns of 43 testicular germ cell tumours were evaluated. CD3 + and CD45RO + (UCHL1 +) T lymphocytes, CD68 + and MAC 387 + macrophages, CD20 + (L26 +) B lymphocytes, and kappa and lambda + plasma cells were counted. The number of immunocompetent cells per mm² of tumour tissue, excluding the necrotic areas, was evaluated. Microscopic fields were randomly selected by two observers. In order to guarantee randomization each surface was divided into parts, numbered through a lattice, and some fields were chosen via a random numbers table. This procedure yielded significantly different counts from those obtained on subjective selection. The number of T-lymphocytes and macrophages was higher in seminomas than in the non-seminomatous testicular germ cell tumours (P < 0.05) Embryonal carcinomas had more T-lymphocytes than immature teratomas. No significant differences were found among testicular germ cell tumours with regards to the B-lymphocytes, with the exception of the high number of B-lymphocytes in mature teratomas. Kappa + and lambda + plasma cells were few in the testicular germ cell tumours. Randomization in the quantification of immunocompetent cells in testicular germ cell tumours is a good means for evaluation of immune response in all the tumour mass, not only in the areas with the most intense inflammatory cell infiltrate, and permits comparison of testicular germ cell tumours with other malignant tumours. Study of immunocompetent cells in every histological type of testicular germ cell tumour is useful in comparing them with other extra-testicular germ cell tumours.     

Histological and histopathological studies on the protective role of Echinacea purpurea extract after intra-testicular injection of magnetic nanoparticles in male albino rats

Local intra-testicular injection for drug release is an important methodology for testicular diseases therapy. Magnetic nanoparticles (MNPs) are new nanomaterials which have several medical applications. This study aimed at investigating the time-dependent toxicity of MNPs after intra-testicular injection, and the protective effect of Echinacea purpurea (EP) extract as antioxidant and immune cells activator. To investigate the protective role of EP extract against MNPs toxicity in testicular tissues, EP extract was simultaneously administrated with MNPs. The intra-testicular injection of MNPs caused spermatogenic apoptosis, cellular necrosis, and interstitial fibrosis. Also, MNPs were found freely in the interstitial area attached to fiber bundles and to lamina propria of the seminiferous tubules. Simultaneous EP extract administration with MNPs injection for 11 wk reduced MNPs toxicity, compared to MNPs only. Also, most MNPs were co-localized in some interstitial cells/macrophages after this EP extract simultaneous administration. In this regard, MNPs were found as groups aggregated in lysosomal vacuoles of macrophages after EP extract administration. In the case of MNPs only injection, MNPs were found freely outside interstitial cells, attached to fiber bundles and surrounding myoid cells of seminiferous tubules. To conclude the simultaneous administration of EP extract/MNPs injection changed both cellular and subcellular biodistribution of MNPs in the testicular tissues. Our present findings indicate a prominent protective role for EP extract as an antioxidant and immune activator against MNPs intra-testicular injection-induced toxicity and may promise a new strategy for testicular disease therapies using nanomaterials.     

Spermiophages on testicular fine needle aspiration cytology: A rare finding

Macrophages usually reside in the testicular interstitial tissues and are normally not found within the seminiferous tubules. However, in certain cases of male infertility, the macrophages are activated and can then be found within the tubules where they can ingest spermatozoa and are labeled as “spermiophages.” FNAC was performed in a 36 year male with history of primary infertility. On microscopy, smears made from right testis were indicative of hypospermatogenesis. On the contrary, smears made from the left testis were very cellular showing Sertoli cells and the entire spectrum of normal spermatogenesis. Also seen were many isolated spermiophages. The cytological impression given for the left testis was normal spermatogenesis with numerous spermiophages. Thus the patient fell in the category of obstructive azoospermia (OA). According to currently adopted hypothesis, macrophages carry ingested sperm heads with some antigenic components to the basal capillaries which may result in the formation of autoantibodies against the spermatozoa. This situation may further diminish the chances of fertility in men. The origin of these spermiophage cells is unknown. Although commonly reported in semen and epididymal biopsies, they have not been reported to occur on testicular fine needle aspiration cytology (FNAC). In our case, no sperms were found on semen examination which were easily picked up on testicular FNAC indicating usefulness of the latter in the diagnosis of cases of male infertility and eliminating the need for a testicular biopsy. Diagn. Cytopathol. 2016;44:232–234. © 2015 Wiley Periodicals, Inc.     

In situ observation of inflammatory cell-tumor cell interaction in human seminomas (germinomas): light, electron microscopic, and immunohistochemical study.

The precise functional significance of the inflammatory cells that infiltrate seminomas remains poorly understood. The present study analyzed 15 cases of testicular and extragonadal seminomas (germinomas) by light and electron microscopy, as well as by immunohistochemical methods, with emphasis on the inflammatory cell-tumor cell interaction. Ultrastructurally, in all 15 cases the lymphocytes (mainly consisting of small lymphocytes) were found to be in intimate contact with the intact tumor cells and with those that displayed damage of varying degree. In particular, relatively early damage, such as local loss of the membrane and/or cytoplasm, occurred at the contact regions. Often, the lymphocytes penetrated deeply into the cytoplasm, even into the nucleus of the tumor cell. In spite of the severe damage to the tumor cells, the lymphocytes were themselves intact. The stromal cells contacted by lymphocytes did not show damage. The tumor cells were in contact with epithelioid cells of granulomas in six cases and scattered macrophages in 11 cases showed damage similar to that seen in tumor cells in contact with lymphocytes. The great majority of the lymphocytes were UCHL1-positive cells. L26- or Leu-7-positive cells were rarely found. The epithelioid cells and scattered macrophages were positive for MAC387. The present morphologic study suggests that the infiltrating lymphocytes, epithelioid cells (probably derived from macrophages), and macrophages may be directly cytotoxic to the tumor cells in the microenvironments of testicular and extragonadal seminomas (germinomas).     

Characteristics of testicular lesions in mice infected with a low dose of encephalomyocarditis (EMC) virus

We investigated the characteristics of testicular lesions induced in mice with a low dose (10 plaque forming units/mouse) of the D variant of encephalomyocarditis (EMC-D) virus. The virus titers of blood and testis peaked at 5 days post-inoculation (5 DPI) and were no longer detected at 14 DPI. The IFN-gamma and iNOS mRNAs expression in the testis and spleen detected by RT-PCR was prominently elevated at 7 DPI, although the expression level of TNF-alpha mRNA was not affected. Signals of viral RNA were clearly detected in degenerative germinal epithelia (in situ hybridization) at 7 DPI, which were surrounded by a small number of macrophages and a few CD4 + T cells and CD8 + T cells (immunohistochemistry). Signals were no longer detected at 21 DPI when seminiferous tubules were highly degenerative and accompanied with infiltration of many macrophages and a small numbers of CD4 + T cells and CD8 + T cells. At 35 DPI, marked atrophy of germinal epithelia composed of Sertoli cells alone was observed, and there were almost no infiltrating cells detected. The present results suggest that macrophages may play an important role in the development of testicular lesions induced in mice with a low dose of EMC-D.     

Requirement of B Lymphocytes in Local Adoptive Transfer of Experimental Allergic Orchitis (EAO) by Lymph Node Cells

ABSTRACT: The necessity of T lymphocytes in local adoptive transfer of experimental allergic orchitis (EAO) is an established fact. The present study was undertaken to elucidate the participation of B lymphocytes in the local transfer system of EAO. The capacity of purified lymph node T and B cells to transfer EAO was compared. Donor strain 13 guinea pigs were sensitized with testicular antigen and complete Freund's adjuvant. We then injected 1 × 10⁸ cells just under the tunica albuginea of the testis of a syngeneic recipient. The recipients were killed 5 days after cell transfer. Nylon-wool nonadherent T cells alone produced only minimal lesions, but could induce more severe mononuclear lesions with the addition of syngeneic, normal macrophages. In contrast, B-cell enriched populations either alone or combined with a small number of macrophages, were able to transfer extensive lesions. In the combined transfer of immune T and immune B cells the severity of transferred lesions was dependent upon the number of B cells. These results suggest that cooperative interactions among T cells, B cells, and macrophages are involved in the generation of transferred testicular lesions, with emphasis on the participation of B cells.     

A method for detecting cytophilic activity in a homologous system

A passive direct technique was developed in order to demonstrate cytophilic activity against homologous spermatozoa is sera of guinea pigs immunized with homologous testicular antigen. A natural adherence between spermatozoa and normal macrophages, called natural cytophilic activity, was found. Specificity of the technique was established since high levels of cytophilic activity were present when each antiserum was made to react with the corresponding antigen. Peritoneal cells other than macrophages did not attach sperm cells to their surface.