线探针核酸杂交分析贵州地区HCV感染株基因型

目的　分析贵州地区丙型肝炎病毒（ＨＣＶ）感染株的基因型。方法　用第二代线探针核酸杂交方法,对９８ 例ＨＣＶ感染者血清进行基因分型。结果　１２ 例献血员血清,均为ＨＣＶ１ｂ 型;１５ 例血液病患者中,１４ 例（９３－３％ ）为ＨＣＶ１ｂ ,１ 例（６－７％ ）为ＨＣＶ１ｂ ＋ ２ 型。慢性丙肝患者３７ 例中,ＨＣＶ１ｂ 型３４例（９１－９％ ） ,混合感染基因型３ 例（ＨＣＶ１ａ ＋ １ｂ 、ＨＣＶ１ｂ ＋ ２ 、ＨＣＶ１ａ ．１ｂ ＋ ２ａ２ｃ 各１ 例）。静脉药瘾者３４ 例,其中ＨＣＶ１ｂ 感染１３ 例（３８－２％ ） ,ＨＣＶ６ａ１０ 例（２９－４％ ） ,ＨＣＶ３ｂ４ 例（１１－８％ ） ,ＨＣＶ３ａ１ 例（２－９ ％） ,混合感染６ 例（１７－６ ％） ,大多为ＨＣＶ２ 复合其他基因型。结论　贵州地区ＨＣＶ感染者中,以ＨＣＶ１ｂ 基因型为主,其次为ＨＣＶ６ａ ,尚存在ＨＣＶ３ａ 和ＨＣＶ３ｂ 。其中ＨＣＶ３ａ、３ｂ 、６ａ ,在本地区系首次报告,但主要存在于静脉药瘾者中。虽然有混合感染基因型的存在,但其基因基础需测序分析     

Human C7 polymorphism: Quantitative analysis of different phenotypes

C7 polymorphism was investigated in a Japanese population (Tokyo). The C7 protein concentrations and hemolytic C7 activities in the serum samples of various phenotypes were measured by radial immuno-diffusion (RID) and radial diffusion hemolysis (RDH). The mean C7 protein levels for the common phenotypes C7 1, C7 2-1, C7 3-1, and C7 4-1 were 88%, 103%, 63%, and 94% of a standard, while the mean C7 activity levels were 96%, 90%, 63%, and 74%, respectively. Both the protein and the activity levels for C7 3-1 were significantly lower than those determined for the most common phenotype C7 1. The mean levels in two individuals of C7 3 phenotype were only 34% in protein concentration and 47% in functional activity. Concerning the ratio of functional to immunochemical C7 (i.e. the relative specific activity), there was no difference between the phenotypes C7 1 and C7 3-1. On the other hand, the mean hemolytic activity and the relative specific activity for C7 4-1 were significantly lower than those for C7 1.     

Laboratory and clinical evaluation of white blood cell differential counts. Comparison of the Coulter VCS, Technicon H-1, and 800-cell manual method

Eight hundred-cell manual white blood cell differential count evaluations of Coulter VCS and Technicon H-1 included estimates of accuracy, imprecision, random and systematic errors, clinical relevance, and detection of immature neutrophils. Accuracy was acceptable, except for H-1 testing of monocytes. Instrument imprecision was similar, except for tighter monocyte values as measured by the VCS. Deming regression determined random errors were greater for H-1 monocytes and proportional errors were similar. Constant errors for the VCS were less than the H-1 for neutrophils and monocytes and greater than the H-1 for eosinophils and lymphocytes. H-1 morphologic false-negative rates were twice those for VCS. True-positive and false-negative rates in cases with immature neutrophils were 70.6% and 20.3% for the VCS and 35% and 55.9% for the H-1. The reference false-positive rate was 4.4%. Clinically appropriate VCS flags were generated in distributionally abnormal cases. Predictive values were higher for the VCS.     

2771例肺肿瘤临床病理特征分析

目的：根据WHO(2015版)肺肿瘤组织学分类标准,探讨军事医学科学院附属医院肺肿瘤的病理类型及分布特点。方法：收集2010年11月1日至2015年3月31日病理诊断2771例肺肿瘤,复习其临床资料、HE切片及免疫组织化学切片。按WHO(2015版)分类标准进行病理诊断及分类。结果：2771例肺肿瘤中多数为男性1671例(60.30%),少数为女性1100例(39.70%);左肺1286例(46.41%)、右肺1456例(52.54%)、双肺29例(1.05%);年龄16~91岁。腺癌1622例(58.53%)、鳞癌424例(15.30%)、腺鳞癌32例(1.15%)、神经内分泌肿瘤465例(16.78%)、大细胞癌19例(0.69%)、梭形细胞癌1例(0.04%)、巨细胞癌1例(0.04%)、癌肉瘤17例(0.61%)、淋巴上皮样癌1例(0.04%)、唾液腺型肿瘤4例(0.14%)、腺瘤11例(0.40%)、间叶性肿瘤55例(1.98%)、淋巴瘤7例(0.25%)、异位起源性肿瘤13例(0.47%)、转移性肿瘤99例(3.57%)。非小细胞肺癌-非特指型,倾向于腺癌小活检中TTF-1抗体阳性率为92.60%(1263/1364)、NapsinA抗体阳性率为97.07%(1324/1364);非小细胞肺癌-非特指型,倾向于鳞癌小活检中CK5/6阳性率为96.99%(290/299)、P40阳性率为98.51%(66/67)、P63阳性率为93.72%(343/366)。非小细胞肺癌分子亚型首次活检中EGFR基因突变率34.77%(378/1087),KRAS基因突变率2.92%(24/823),BRAF基因突变率0.87%(5/572),ALK融合基因阳性率11.99%(41/342),ALK-D5F3抗体阳性率22.23%(66/296),ROS1融合基因阳性率2.56%(7/273),c-Met基因扩增率4.40%(12/273),c-Met基因突变率6.67%(1/15),Her-2基因突变率0.73%(2/273),PIK3CA基因突变率13.33%(2/15),PTEN基因突变率6.67%(1/15),RET融合基因阳性率0(0/15)和NTRK1融合基因阳性率0(0/15)。二次活检率EGFR基因4.76%(18/378),ALK融合基因2.44%(1/41)。结论：肺肿瘤在男性患者中高发,病变部位最常见于右肺,腺癌已经成为最常见的病理类型。首次活检中非小细胞肺癌驱动基因中EGFR基因、ALK融合基因存在较高的突变率,其他基因突变率虽低但不容忽视。二次活检率较低,需引起重视。     

Detection of aneuploidy in human spermatozoa using fluorescencein situ hybridization (FISH)

Summary Fluorescencein situ hybridization (FISH) with chromosome specific alpha-satellite DNA probes was used to estimate the rates of aneuploidy of chromosomes 1, 17, 18, X and Y in human ejaculated sperm. Sperm samples were collected from six donors, and biotinylated DNA probes, D1Z5, D17Z1, D18Z1, DXZ1 and DYZ3 were hybridized to interphase sperm which had been pretreated with dithiothreitol to expand their nuclei. A minimum of 3,000 sperm per donor were analyzed. The hybridization efficiency was 99.68% for all the five probes. The frequencies of aneuploidy for chromosomes 1, 17 and 18 were 0.65%, 0.66% and 0.61% respectively. For XX-and YY-sperm the frequencies were 0.28% and 0.27% respectively. To estimate the diploidy and disomy rates, a mixture of D17Z1 and D18Z1 were used as probes, and the frequency of diploid sperm was calculated to be 0.27%. After subtraction of the diploidy rate, the disomy rates for chromosomes 1, 17, and 18 were estimated to be 0.38%, 0.39% and 0.33%, respectively. The proportion of X- and Y-bearing sperm were 49.9% and 49.66%, consistent with an expected 1: 1 ratio.     

贵州三个少数民族谷胱甘肽S-转移酶M1、T1和P1基因多态性

目的研究贵州省从江县侗族、威宁县彝族、荔波县瑶族的谷胱甘肽S-转移酶基因(GSTs)多态性。方法在隔离自然人群(从江县侗族108人、威宁县彝族104人、荔波县瑶族109人)中,采用多重等位基因特异聚合酶链反应方法分析GSTM1和GSTT1基因多态性,采用聚合酶链反应及限制性片段长度多态性方法分析GSTP11578(A→G)基因多态性。结果贵州省从江县侗族、威宁县彝族、荔波县瑶族的GSTM1和GSTT1纯合缺失基因型频率分别为59.6%～71.2%、39.4%～72.5%。其GSTP11578(A→G)基因型频率分别是:AA为63.3%～75%、AG为23.2%～35.8%、GG为0～1.9%。等位基因频率:A为81.2%～86.6%,G为13.4%～18.8%。结论GSTT1基因型频率在贵州从江侗族、威宁彝族、荔波瑶族中存在差异,其分布特征可能与人群中不同种族以及同一种族不同民族相关。     

北京市甲型H1N1型流感病毒基因检测与流行趋势分析

目的 通过对甲H1N1型流感病毒基因的检测分析,揭示其基因变异情况及探讨甲型流感病毒的流行趋势,为甲型流感的防控提供理论依据.方法 样本为患者鼻咽部的粘膜上皮细胞和分泌物.取200μL标本置于核酸分离纯化系统中提取核酸,然后以LightCycler 480 PCR仪进行核酸扩增,最后分析曲线判断结果.每份标本均对甲型H1N1流感病毒基质蛋白基因（M基因）、核蛋白基因（NP基因）、血凝素基因（HA基因）、人类的RNA酶P基因（RNase P基因）进行检测和分析.结果判定：M基因、NP基因、HA基因、RNase P基因均阳性,为真阳性;HA基因阴性,其它三种基因阳性,为可疑阳性.结果 共检测3673份样本,其中,阴性2404份,甲型流感阳性样本共1269份,阳性率34.5％;阳性样本中季节性甲型流感641份,占50.5％;甲型H1N1流感阳性628份,占49.5％（真阳性433份,占68.9％,可疑阳性195份,占31.1％）.在整个流行季,真阳性与可疑阳性在相同时间的检出率比值基本不变.甲型H1N1检出高峰在10月,季节性甲流则分别在9月和12月.中青年人群的阳性检出率为57.5％,儿童为39.9％,而60岁以上的老年人仅占2.6％.结论 甲型H1N1流感以M、HA、NP、RNaesP四种基因阳性毒株为主,其流行高峰出现在10月份,感染人群以中青年和儿童为主.     

HIV-1 subtypes D and F are prevalent in Guinea Conakry

BackgroundLimited data is available upon the distribution of different HIV-1/2 genotypes in the blood donor population from Guinea Conakry.ObjectivesTo investigate the prevalence of HIV-1/2 subtypes in asymptomatic blood donors in Guinea Conakry, in order to update knowledge of HIV-1/2 epidemiology within this country.Study designSamples from 104 blood donors seropositive for HIV-1/2 were tested for HIV-1 by real-time RT-PCR. Those negative for HIV-1 were tested with HIV-2 nested RT-PCR. Positive samples were further amplified in the HIV-1 gag and pol regions and sequenced. Subtypes were determined by phylogenetic analysis on amplicon sequences.Results61 samples were positive by HIV-1 real-time RT-PCR. Of the 43 negative, 2 (4.6%) were positive for HIV-2. 52/61 (85.3%) samples were positive by nested RT-PCR. Of the 52, 43 (70.5%) and 31(59.6%) sequences were obtained in the gag and pol regions, respectively; 23 for both regions. HIV-1 subtype distribution was 1 B (2.1%), 8 F (17%), 8 D (17%) and 28 CRF02_AG (59.6%) with 2 unclassified recombinants (4.3%). Unique clusters for subtype D and F distinguished Guinea from HIV-1 subtype distribution in neighboring countries.ConclusionsSubtype F and subtype D strains, uncommon in West Africa, are a substantial part of HIV-1 epidemiology in Guinea.     

Immunocytochemical colocalization of fibroblast growth factor-1 with neurotrophin-3 in mouse alveolar macrophages

Alveolar macrophages are known to express a variety of growth factors and neurotrophins. Fibroblast growth factor-1 (FGF-1) is abundantly present in the lung and has mitogenic and neurotrophic activities similarly to neurotrophins. In order to determine whether FGF-1 associates with neurotrophins in alveolar macrophages, we investigated the immunocytochemical colocalization of FGF-1 with neurotrophins, nerve growth factor (NGF), brain-derived neurotrophic factor (BDNF), and neurotrophin-3 (NT-3), in mouse alveolar macrophages. The results showed that 34% of macrophages were immunoreactive for FGF-1, 10% for NGF, 9% for BDNF, and 17% for NT-3. Of FGF-1-immunoreactive (IR) macrophages, 16% were immunoreactive for NT-3, but only small percentages were immunoreactive for NGF (0.8%) and for BDNF (0.3%). FGF-1 and neurotrophins were all localized in the intracellular vesicles. In the vesicles, FGF-1 and NT-3 were frequently colocalized. All macrophages expressed lysosome-associated protein-2 (LAMP-2), a late endosomal and lysosomal marker, and early endosomes antigen 1 (EEA1), an early endosomal marker. FGF-1 and NT-3 were predominantly colocalized with LAMP-2 rather than with EEA1, whereas NGF and BDNF were colocalized with EEA1 rather than with LAMP-2. These results indicate that FGF-1 and NT-3 are substantially expressed in mouse alveolar macrophages and colocalized in vesicles, predominantly in late endosomes and lysosomes.     

HBME-1, MOC-31, WT1 and calretinin: an assessment of recently described markers for mesothelioma and adenocarcinoma

AIMS: To evaluate HBME-1, WT1, calretinin and MOC-31 in the differential diagnosis of pleural mesothelioma and adenocarcinoma of the lung. METHODS AND RESULTS: Paraffin-embedded formalin-fixed blocks from six reactive pleuras, 42 mesotheliomas and 40 adenocarcinomas were used. Sections were stained for Leu-M1, HBME-1, calretinin, WT1 and MOC-31. Leu-M1 was positive or equivocal in 34% of mesotheliomas and in 78% of adenocarcinomas; reactive pleuras were all negative. HBME-1 was positive or equivocal in 76% of mesotheliomas and in 73% of adenocarcinomas; five reactive pleuras were positive. Calretinin was positive or equivocal in 92% of mesotheliomas and in 73% of adenocarcinomas; two reactive pleura were equivocal and four were positive. WT1 was positive or equivocal in 72% of mesotheliomas (excluding autopsy cases) and in 20% of adenocarcinomas; all reactive pleuras were positive. MOC-31 was positive or equivocal in 5% of mesotheliomas and in 90% of adenocarcinomas; all reactive pleuras were negative. The reaction with Leu-M1 was graded as equivocal in 25% of the adenocarcinomas. All 24 of the autopsy cases of mesothelioma were negative for WT1 and in many operative specimens only the periphery was stained. CONCLUSIONS: Neither calretinin nor HBME-1 are sufficiently discriminatory to be of use, even as members of a panel of antibodies. WT1 shows some promise, but it cannot be used on autopsy material. The utility of MOC-31 is confirmed, and outperforms Leu-M1.     

Oligotyping of Italian celiac patients with the 11th International Histocompatibility Workshop reagents.

Oligotyping for HLA-DRB1, DQA1 and DQB1 specificities has been performed on PCR-amplified DNA from 55 Italian celiac children, living in Rome, and 50 blood donors. 52.6% of CD patients were DR3;DQA1*0501;DQB1*0201-positive versus 14% of controls (RR = 6.85) and 34.5% were DR5,7;DQA1*DQB1*0201-positive versus 2% of controls (RR = 25.86). 7 patients (12.7%) were negative for the DQA1*0501/B1*0201 dimer: 3 of them were DR4 (5.4%) and the others typed as DR1,5; 1,7; 5,7 and w6,7. No patient was negative for both DQA1*0501 and DQB1*0201 alleles.     

Prevalence of HIV-1 non-B subtypes, syphilis, HTLV, and hepatitis B and C viruses among immigrant sex workers in Madrid, Spain

Sexually transmitted disease (STD) remains a major public health challenge in developed countries, exacerbated by the advent of the HIV epidemic. The objectives of this study were to assess the prevalence of serological markers of syphilis, HIV-1/2, HTLV-I/II, HBV, and HCV infections among immigrant sex workers in Madrid, Spain and to characterize the HIV-1 variants in seropositive individuals. Sera from 762 immigrant commercial sex workers (75.3% from sub-Saharan Africa, 18.2% from South America, and 6.4% from Eastern Europe) were collected between 1998 and 2003 in Madrid and examined. Antibody detection was performed by screening assays (RPR, ELISAs) and confirmed by FTA-Abs, LIAs and Western-blot tests. HIV-1 subtyping was carried out by phylogenetic analyses of the protease and envelope genes. Antibodies to HIV-1 were found in 5.2%, while 3.5% tested positive for HBsAg, 3% for syphilis antibodies, 0.8% for HCV antibodies, and 0.2% for HTLV-I antibodies. None were reactive for HIV-2 or HTLV-II antibodies. HIV-1 seroprevalence among Africans and Ecuadorians was 4.5 and 10.9%, respectively. All HIV-1 seropositive Ecuadorians were transsexual men, and 28.6% had active syphilis infection. Up to 80% of HIV-1 positive specimens were characterized as non-B subtypes, with subtypes G, A, and G/A recombinants being the most frequent among African individuals. In contrast, South Americans with HIV-1 infection carried exclusively subtype B variants. A relatively high proportion of immigrant sex workers in Madrid were infected with HIV-1 and syphilis, whereas infections with hepatitis viruses or HTLV were uncommon.     

Role of GPER1, EGFR and CXCR1 in differentiating between malignant follicular thyroid carcinoma and benign follicular thyroid adenoma

It is extremely difficult to discriminate between follicular thyroid carcinoma (FTC) and follicular thyroid adenoma (FTA) before surgery, because the morphologies of carcinoma cells and adenoma cells obtained by fine needle aspiration biopsy (FNAB) are similar. Molecular markers may be helpful on this issue. The purpose of this study was to assess the role of GPER1, EGFR and CXCR1 in differential diagnosis between FTC and FTA. GPER1, EGFR and CXCR1 mRNA expression levels were examined in 15 FTCs and 10 FTAs using real-time RT-PCR. FTC showed to have significantly increased mRNA levels of the three molecules compared to FTA (P < 0.001 for all the three molecules). GPER1, EGFR and CXCR1 protein expression in 106 FTCs and 128 FTAs were analyzed using immunohistochemistry. The rates of GPER1, EGFR and CXCR1 high expression were 73.6%, 72.6% and 70.8% in FTC and 30.5%, 28.1% and 27.3% in FTA, respectively. Statistical analysis showed that GPER1, EGFR and CXCR1 protein expression were correlated with one another in FTC and concomitant high expression of the three molecules had stronger correlation with the occurrence of FTC than did each alone. The positive predictive values (PPV) for concomitant high expression of the three molecules for discriminating between FTC and FTA were 91.0% for GPER1/EGFR, 93.8% for GPER1/CXCR1, 92.3% for EGFR/CXCR1 and 98.2% for GPER1/EGFR/CXCR1, respectively. These results indicated that the evaluation of GPER1, EGFR and CXCR1 concomitant high expression may be helpful in differential diagnosis between FTC and FTA.     

402例甲型H1N1流感病例流行病学特征分析

目的了解深圳市福田区甲型H1N1流感病例的流行病学特征,为甲型H1N1流感预防控制提供科学依据。方法应用描述性流行病学方法分析402例甲型H1N1流感病例的流行病学特征,包括病例的年龄、性别、职业分布、地区分布等。结果 402例甲型H1N1流感病例男性占58.71%,女性占41.29%;90.55%的病例集中在0～30岁年龄段,11～20岁年龄段病例最多,占总病例数的41.79%;从职业构成看,所有病例中,绝大多数为学生,占总病例数的64.18%。结论学生是甲型H1N1流感的危险人群,应当注重学校等场所的甲型H1N1流感防控工作。     

汕头市濠江区甲型H1N1流感病毒感染状况

目的了解汕头市濠江区甲型H1N1流感感染状况,为防控措施提供科学依据。方法在2010年1-9月期间,开展3次人群甲型H1N1流感病毒感染状况横断面调查,采用血凝抑制法检测居民甲型H1N1流感病毒血清抗体,对结果进行统计分析。结果 3次人群甲型H1N1流感病毒感染血清学检测,共检测480人,总阳性率为15%,各月份阳性率逐次下降（1月为20%、3月为15.2%、9月为9.4%）;各年龄组人群中,6～岁组阳性率（30.2%）最高,16～岁组阳性率（15.1%）次之,0～岁组幼儿阳性率（8.3%）再次之,≥60岁老人组阳性率（6.3%）最低。结论濠江区人群甲型H1N1流感病毒感染率较低,人群免疫屏障尚未完全建立,适当接种甲流疫苗仍有必要。     

Treatment outcome and relapse with short-term oral terbinafine (250 mg/day) in tinea pedis.

A total of 168 patients with tinea pedis, but without onychomycosis, were treated with 1 cycle of terbinafine (TBF) (1 cycle: defined as 250 mg/day for 1 week). KOH preparation for direct microscopy was performed 4, 8 and 12 weeks after starting therapy to determine if testing was positive for tinea. Patients with no negative results on KOH examination or no evidence of obvious clinical improvement at 8 weeks, another cycle of the therapy was prescribed. The "cure", "no cure", "dropout", and "discontinuation/unevaluable" rates were 89.3%, 4.8%, 4.8% and 1.2%, respectively. The number of cycles required for cure in the plantar type was 1 cycle in 65.9% and 2 cycles in 54.5% of cases; in the interdigital type, 1 cycle in 79.1% and 2 cycles in 20.9% of cases; and mixed type, 1 cycle in 29.1% and 2 cycles in 60.9% cases. Among patients who were followed for at least 3 years after cure, the relapse rates were about 10% each year: 1 year, 11.3%; 2 years, 8.9%; and 3 years, 11.2%. The relapse rate of about 10% each year over a 3-year period suggests that reinfection may be likely.     

Value of PAX8 and WT1 Immunostaining in Confirming the Ovarian Origin of Metastatic Carcinoma in Serous Effusion Specimens

We evaluated the detection rates of PAX8 and WT1 immunostaining in 68 (45 as cell blocks, 23 as smears) serous effusion specimens that had a cytologic diagnosis of metastatic carcinoma of ovarian origin. Of the cases, 58 (85%) were positive for PAX8, 56 (82%) were positive for WT1, and 64 (94%) were immunoreactive with either or both markers. Detection rates of PAX8 and WT1 were 85% (44/52) and 92% (48/52), respectively, for metastatic serous carcinoma and 100% (5/5) and 20% (1/5), respectively, for metastatic clear cell carcinoma. Detection rates using cell blocks and smears were 91% and 78%, respectively, with PAX8 and 82% and 83%, respectively, with WT1. We concluded that PAX8 and WT1 had comparable overall detection rates in confirming ovarian origin of malignant effusion. The combination of both markers substantially improved the detection rate. Cell blocks and smears can be used for staining, but a cell block is preferred for PAX8 staining.     

Aberrant keratin expression is common in primary hepatic malignant vascular tumors: A potential diagnostic pitfall

Malignant vascular neoplasms such as epithelioid hemangioendothelioma (EHE) and angiosarcoma (AS) can arise within the liver. The aim of this study was to study the expression of keratins CK7, AE1/AE3 and OSCAR in primary hepatic EHE and AS. 9 cases of hepatic EHE and 13 cases of hepatic AS were stained with ERG, CK7, keratin AE1/AE3 and keratin OSCAR. Their expression was graded as 1+ (1–25% of tumor cells positive), 2+ (26–50%), 3+ (51–75%) or 4+ (>75%). ERG was positive in all 9 (100%) EHEs and all 13 (100%) ASs. CK7 was positive in 5/9 (56%) EHEs (2, 1+; 1, 2+; 1, 3+; 1, 4+) and 1/13 (8%) AS (2+). Keratin OSCAR was positive in 6/9 (67%) EHEs (5, 1+; 1, 2+) and 4/13 (31%) ASs (2, 1+; 1, 2+; 1, 4+). Keratin AE1/AE3 was positive in 6/9 (67%) EHEs (3, 1+; 3; 2+) and 4/13 (31%) ASs (2, 1+; 1, 2+; 1, 4+). Overall, 6/ 9 (67%) EHEs were positive for at least one keratin marker, of which 5 were positive for all 3 keratins (AE1/AE3, OSCAR and CK7) while 1 was positive only for 2 keratins (OSCAR and AE1/AE3). 4/13 (31%) of ASs were positive for both keratins OSCAR and AE1/AE3, of which 1 case was also positive for CK7. Aberrant keratin expression is common in primary hepatic EHEs (67%) and ASs (31%). Awareness of this diagnostic pitfall is important for avoiding misdiagnosis of these primary hepatic malignant vascular tumors as carcinomas.     

Immunoreactivity for IL-1 beta and TNF alpha in human lymphoid and nonlymphoid tissues.

Monoclonal antibodies (MAbs) against two non-cross-reacting antigens of human IL-1 beta (Vhp20 and BRhC3) and human TNF alpha (B154.2 and B154.7) were applied to identify cytokine-containing cells in tissue sections and in cell suspensions. IL-1 beta- or TNF alpha-positive cells were not present in immunostained cytocentrifuge smears prepared from freshly isolated peripheral blood leukocytes, spleen, and lymph node cells. After 18 hours of culture with bacterial endotoxin (LPS), 80% to 90% of blood monocytes, 30% of spleen macrophages, and 2% to 28% of lymph node macrophages were strongly positive for IL-1 beta with either of the MAbs. Furthermore, 25% to 35% of blood monocytes and 6% to 60% of lymph node macrophages were stained for TNF alpha. Cells positive for IL-1 beta or TNF alpha were extremely rare in sections of normal thymus, spleen, and lymph nodes. Immunoreactivity for IL-1 beta or TNF alpha was frequently observed in sections of granulomatous lymphadenitis (N = 11). IL-1 beta or TNF alpha staining was confined to the epithelioid macrophages forming the granuloma, and the intensity of TNF alpha reactivity was generally stronger. The high frequency of cytokine-containing cells in this pathologic condition was confirmed in a cell suspension study showing that 20% of epithelioid macrophages were weakly positive for IL-1 beta and 80% were strongly positive for TNF alpha. The presence of cytokine-containing cells was investigated in cryostat sections of several nonlymphoid organs with normal histologic appearance. IL-1 beta reactivity was not observed in any of the tissues. TNF alpha reactivity was frequently demonstrated in isolated macrophages embedded in the interstitial connective tissue.