Distribution and immunophenotype of the inflammatory cell population in the benign lymphoepithelial lesion (Mikulicz's disease)

Benign lymphoepithelial lesion (BLL) is an autoimmune process characterized by swelling and diffuse inflammation of the major salivary glands. Autoantibodies have been isolated from lymphocyte cultures obtained from affected salivary glands, but the pathogenesis is still unknown. Previous studies have shown that the predominant population of inflammatory cells is represented by helper T cells, with only brief mention of the B cell population. Twenty-five surgical specimens from patients with BLL were studied immunohistochemically. Antisera used included monoclonal antibodies LN-1 and LN-2 for B cells, LN-3 for cells expressing human leukocyte antigen-DR (HLA-DR) antigens, UCHL-1 for T cells, Leu-7 for natural killer (NK) cells, and T suppressor lymphocytes and the polyclonal antibody to S100 protein for dendritic cells. A peculiar distribution of the inflammatory infiltrate was observed in all cases, characterized by the presence of very irregular "germinal centers" with pseudopod-like extensions surrounding epimyoepithelial islands. Lymphoid cells in this location were reactive with LN-1 and LN-2 antibodies. These structures were surrounded by a "mantle" of mixed small B and T lymphocytes. A well-defined "interfollicular" zone was composed of cells strongly reactive with UCHL-1 and LN-3 antibodies, indicating the presence and activation of T cells. Dendritic cells defined by S100 and LN-2 reactivity were intermixed with epimyoepithelial cells, and were identified in 18 cases. Epithelial expression of HLA-DR antigens was restricted to inflamed areas. In contrast to previous reports denying the presence of Leu-7-positive cells in these lesions, cells reactive for this antibody were identified in 13 of 20 cases, predominantly within germinal centers. The presence of dendritic cells, complex organization of the inflammatory infiltrate into well-defined B cell proliferation centers and activated interfollicular T areas, and the abnormal expression of HLA-DR antigens in epithelial cells support an antibody-mediated destruction of the epithelial cells in this disease.     

Pituitary adenoma,primary parathyroid hyperplasia and papillary (non-medullary) thyroid carcinoma

Summary An acidophilic pituitary adenoma associated with primary nodular parathyroid hyperplasia and a small papillary thyroid carcinoma was discovered at the autopsy of a 44 year old female acromegalic. The thyroid carcinoma showed evidence of lymphatic spread. Several etiopathogenetic mechanisms for the non-medullary thyroid carcinomata associated with Multiple Endocrine Neoplasia (MEN) have been postulated, since the follicular epithelium of the thyroid does not belong to the neural ectoderm derivates unlike the C-cells of the thyroid, the adenohypophysis and probably the parathyroid glands. Apart from genetic influence, or coincidence, one has to rule out carcinogenic exposure or hormonal influence. Clinically speaking, one should always consider whether malignant thyroid disease coexists with hyperplastic or neoplastic parathyroid tissue.     

The spectrum of histologic changes in thyroid hyperplasia: a clinicopathologic study of 300 cases

Thyroid hyperplasia is a physiologic response of follicular epithelium to hormonal changes that result in disturbances in the feedback mechanism of thyrotropin-releasing hormone and thyroid-stimulating hormone. The most common manifestation of this process is the so-called sporadic goiter (diffuse or nodular hyperplasia), a condition that may be associated with a variety of stimuli. Some of the histologic changes that can be observed in hyperplasia of the thyroid can sometimes lead to an incorrect diagnosis of malignancy. We studied 300 consecutive cases of hyperplasia of the thyroid to evaluate morphologic features that could potentially be mistaken for neoplastic conditions. Florid papillary hyperplasia of follicular epithelium was observed in 13% of cases, in several instances closely resembling the papillary structures of papillary thyroid carcinoma. Foci displaying nuclear clearing closely resembling "Orphan-Annie" nuclei were present in 15% of cases. Nuclear grooves and pseudonuclear inclusions were also identified focally in 8% of cases. Cytologic atypia was observed in 7% of cases, including nuclear enlargement, multinucleation, and nuclear pleomorphism with prominent nucleoli. Mitoses were observed in 6% of cases and averaged 1 to 2 per 20 high-power fields. This finding was usually seen in the more cellular areas in cases characterized by a solid, microfollicular pattern of growth. Psammoma bodies were observed focally in 4 cases (1.3%). Infiltration of adjacent skeletal muscle by benign hyperplastic follicles was seen in 3 cases (1%). Another unexpected finding in 2 cases was the identification of small clusters of normal thyroid follicles within the sinuses of lymph nodes located adjacent to the gland (>1%). The present study confirms that thyroid hyperplasia can sometimes display features that may be confused for a malignant neoplastic process. Awareness of such features and their recognition are of importance to avoid a misdiagnosis of malignancy.     

Identification of apoptotic proteins in thyroid gland from patients with Graves' disease and Hashimoto's thyroiditis

Apoptosis, i.e. natural programmed cell death, is a physiological phenomenon indispensable for normal functioning of the organism. The signal to apoptosis can be started practically in any cell. Disturbances in the apoptosis regulation determine the essential link of the pathogenesis of many diseases, including autoimmune thyroid disorders.

The aim of the study was to assess the expression of Fas/FasL and caspase eight in the tissues of the thyroid gland in patients with Graves' disease (GD), non-toxic nodular goiter (NTNG) and Hashimoto's thyroiditis (HT). The analysis of Fas/FasL expression was performed by western blot and immunohistochemical investigation with DAB-visualization and Mayer's hematoxylin staining. Caspase-8 expression in thyroid follicular cells was assayed by western blot method.

Identification of the proapoptotic proteins FasL and Fas exhibited their pronounced expression in the thyroid tissue in GD patients (++; ++) and HT (+++; +++) as compared to the NTNG group (0/+; 0/+). Among the study groups, the expression of caspase-8 was revealed in band 55 kDa from patients with autoimmune thyroid diseases.

In GD patients, the percentage of thyrocytes with FasL expression correlated positively with TRAb (R = 0.58, p < 0.02). However, no such correlations were noted in HT or non-toxic multinodular goiter. There were no significant correlations between thyroid hormones and the percentage of thyrocytes with Fas and FasL expression.

In conclusion, our findings suggest that the changes in the expression of apoptotic molecules on the surface of T lymphocytes and thyroid follicular cells in patients with autoimmune thyroid disorders reflect their substantial involvement in the pathogenesis of GD and HT. In addition, analysis of Fas/FasL and caspase-8 expression in thyroid tissue may indicate the disease activity and immunological phenotype.     

The expression and distribution of S-100 protein and CD 83 in thyroid tissues of autoimmune thyroid diseases

To investigate the expression and distribution of S-100 protein and CD83 in the thyroid tissues of autoimmunethyroid diseases(ATDs),and to study the role of the dendritic cells in the pathogenesis of ATDs,immunohistochemical staining was used on pathological tissues of 20 patients with Hashimoto's thyroiditis(HT)and 20 patients with Graves' disease(GD) to check the expression and distribution of S-100 protein and CD83.Compared with control group(20 cases of thyroid follicular adenoma,TFA),the higher expressions of S-100 inHT(139.38±5.92 vs 59.47±11.69) and GD(119.42±14.48 vs 59.47±11.69) were observed respectively(p<0.001).The increased positive expressions of CD83 which is known as a marker of mature and activated DCs inHT(22.58±13.96 vs 5.19±8.08) and GD(29.92±14.43 vs 5.19±8.08) were also found respectively(p<0.001).Serum TPO antibody(TPO-Ab,67.3±11.6%) and Tg antibody(Tg-Ab,59.8±10.1%) in HT were higher thanthose in GD(28.4±5.7%,23.1±4.9%) and TFA(6.1±3.4%,7.2±4.6%)(p<0.01).Serum TR-Ab in GD(16.3±5.6 U/L) was higher than those in HT(4.8±2.3 U/L) and TFA(2.5±1.2 U/L)(p<0.01).Our findings suggestthat the high expression of DCs' markers may be related to the pathogenesis of HT and GD.The upregulationof both the number and the matured functions of DCs,may lead to present more antigens and to produce moreauto-antibodies(such as Tg-Ab and TPO-Ab in HT,TR-Ab in GD),which may be involved in pathogenesis ofthe autoimmune thyroid diseases.Cellular & Molecular Immunology.2004;1(5):378-382.     

Analysis of MHC genes in a Tunisian isolate with autoimmune thyroid diseases: implication of TNF -308 gene polymorphism

Autoimmune thyroid diseases (AITDs), which include Hashimoto thyroiditis (HT), Graves' disease (GD) and primary idiopathic myxoedema (PIM), are recognized as multifactorial diseases. In this study, we have examined single and haplotypic genetic variation across the major histocompatibility complex (MHC) in a Tunisian isolate with a high prevalence of AITDs (62 patients: 32 with GD, 9 with HT and 21 with PIM). Genotyping was performed for HLA class I and II alleles as well as polymorphisms within tumor necrosis factor (TNF), lymphotoxin alpha (TLalpha) and heat shock protein (HSP70-02 and HSP70-hom) genes. Our results showed association of HLA-A2-B50-TNF 2 haplotype with AITDs (p = 0.045). Linkage analysis using Simwalk2 program has shown significant result with TNF -308 gene polymorphism (p = 0.03). The FBAT has given evidence for genetic association with TNF -308 and HLA-DR gene polymorphisms. TNF 2 allele was associated with GD (p = 0.0011), whereas TNF 1, HLA-DR11 and DR12 (p = 0.0039, p = 0.00089 and p = 0.0056, respectively) were rather implicated in HT pathogenesis. Results found by TDT-STDT have confirmed the involvement of the TNF -308 gene polymorphism in AITD pathogenesis (p < 10(-9)).     

Hepatic lesions in patients treated with synthetic anabolic steriods.

Hepatic abnormalities are described in three patients who received synthetic anabolic steroids. A child with Franconi's anaemia was treated for four years and at necropsy the liver showed generalized hyperplasia, hyperplastic nodules, and a benign hepatoma. Two adults received only three months' therapy with synthetic androgens; in one there was generalized hepatic hyperplasia and in the other widespread nodular hyperplasia. It is suggested that anabolic steroids may induce tumours through intermediate hyperplastic lesions, a sequence similar to that seen during tumour induction by carcinogens in experimental animals.     

Single-cell analysis of intrathyroidal lymphocytes shows differential cytokine expression in Hashimoto's and Graves' disease

Most human organ-specific autoimmune diseases such as Hashimoto's thyroiditis (HT) are considered to be Th1 mediated, and a quantitative dominance of Th1 cells in thyroid infiltrates from both Graves' disease (GD) and HT affected glands has been reported. However, Th2 dominance would be expected in GD, where thyroid hyperfunction induced by stimulating antibodies predominates over tissue destruction. We have analyzed the interleukin-4 (IL-4), interferon-γ (IFN-γ) production by T cells at the single-cell level, both in infiltrating lymphocytes isolated from digested GD and HT thyroid glands and in derived T cell lines, by direct intracellular cytokine detection. Results showed a heterogeneous pattern of cytokine production in bulk GD infiltrates and derived T cell lines, and a similar pattern was observed in the much larger HT infiltrates. Both type 1 and type 2 cytokines were simultaneously produced by the infiltrating populations, and T cells with both patterns as well as intermediate patterns similar to ThO cells could be detected ex vivo. However, the larger T lymphocytes, presumably activated and responsible for the autoimmune damage, predominantly produced IL-4 in GD and IFN-γ in HT. The specificity of the Th2 responses in GD was suggested by the enrichment in IL-4 production after antigen-specific expansion of two oligoclonal T cell lines. These data show that both type 1 and type 2 cytokines are produced in the thyroid glands affected by autoimmunity and that the difference between diseases may be the effect of a functionally dominant population at a given time. This in vivo chronically activated antigen-specific population, producing type 1 or type 2 cytokines locally, may be responsible for the effect finally leading to one of the disease states.     

Immunogenetics of autoimmune thyroid diseases: A comprehensive review

Both environmental and genetic triggers factor into the etiology of autoimmune thyroid disease (AITD), including Graves' disease (GD) and Hashimoto's thyroiditis (HT). Although the exact pathogenesis and causative interaction between environment and genes are unknown, GD and HT share similar immune-mediated mechanisms of disease. They both are characterized by the production of thyroid autoantibodies and by thyroidal lymphocytic infiltration, despite being clinically distinct entities with thyrotoxicosis in GD and hypothyroidism in HT. Family and population studies confirm the strong genetic influence and inheritability in the development of AITD. AITD susceptibility genes can be categorized as either thyroid specific (Tg, TSHR) or immune-modulating (FOXP3, CD25, CD40, CTLA-4, HLA), with HLA-DR3 carrying the highest risk. Of the AITD susceptibility genes, FOXP3 and CD25 play critical roles in the establishment of peripheral tolerance while CD40, CTLA-4, and the HLA genes are pivotal for T lymphocyte activation and antigen presentation. Polymorphisms in these immune-modulating genes, in particular, significantly contribute to the predisposition for GD, HT and, unsurprisingly, other autoimmune diseases. Emerging evidence suggests that single nucleotide polymorphisms (SNPs) in the immunoregulatory genes may functionally hinder the proper development of central and peripheral tolerance and alter T cell interactions with antigen presenting cells (APCs) in the immunological synapse. Thus, susceptibility genes for AITD contribute directly to the key mechanism underlying the development of organ-specific autoimmunity, namely the breakdown in self-tolerance. Here we review the major immune-modulating genes that are associated with AITD and their potential functional effects on thyroidal immune dysregulation.     

p27蛋白在结节性甲状腺肿和甲状腺癌中的表达及意义

目的探讨p27蛋白在结节性甲状腺肿和甲状腺癌中的变化,为研究甲状腺癌的发生机制提供参考依据。方法应用免疫组化法检测结节性甲状腺肿（61例）、结节性甲状腺肿伴增生结节（16例）、甲状腺癌（45例）、正常甲状腺组织（20例）标本中p27蛋白的表达。结果 p27蛋白表达阳性率在正常甲状腺组织为60.5%,结节性甲状腺肿为36.10%,甲状腺癌为33.3%,结节性甲状腺肿伴增生结节为0。可见,结节性甲状腺肿和甲状腺癌的p27蛋白表达阳性率与结节性甲状腺肿伴增生结节相比,差异均有统计学意义（P〈0.01）。结论结节性甲状腺肿可能通过增生结节向甲状腺癌演变。     

Reciprocal/dichotomic expression of vimentin and B cell differentiation antigens in Reed-Sternberg's cells

Summary An immunohistochemical study of 63 cases of Hodgkin's disease was undertaken using formalin-fixed paraffin embedded tissue sections. The antibodies used were against L26, LN-1, LN-2, EMA (epithelial membrane antigen), Leu-M1, Vimentin, UCHL-1, S-100, and lysozyme. Hodgkin's disease could be divided into three groups: the first group was LN-1+/L26+/vimentin-, the second LN-1-/L26+/vimentin+, and the third LN-1-/L26-/vimentin+). Sixteen cases of follicular lymphomas were also examined and were all positive for LN-1 and L26 and negative for vimentin. Thus the vimentin negativity of the first group, including 7 nodular lymphocyte-predominant cases, gives further evidence of their germinal center B-cell origin. Since vimentin is expressed mainly in the immature stage of B-lymphocytes, the second group of Hodgkin's disease may represent immature B-cell Hodgkin's disease. In the third group, vimentin was present in Reed-Sternberg's (RS) and Hodgkin's (H) cells in 45 of the 48 cases (92.5%). In none of 48 cases were these cells positive for S-100 or lysozyme, but strong vimentin-positivity still suggested monocytic or histiocytic origin. The results of our study suggest, at least, divergent origin of RS's and H's cells.     

Expression of vascular adhesion molecules on human endothelia in autoimmune thyroid disorders.

Cellular activation and expression of certain adhesion molecules within vascular endothelium is a critical event in leucocyte recruitment and emigration. A wide array of different adhesion receptors has been identified to mediate the interaction between endothelial cells (EC) and leucocyte subpopulations. In this study, the tissue expression of E-selectin, P-selectin, CD31, and endoglin endothelial cell adhesion molecules was studied on thyroid tissue from patients with Graves' disease (GD) and Hashimoto's thyroiditis (HT). We found an up-regulated expression of E-selectin in EC in GD and HT thyroids, specifically in those areas more severely inflamed, with no reactivity in control thyroids. P-selectin was basally expressed in postcapillary venules in control glands, with an increased expression in HT and GD glands. On the other hand, increased CD31 expression was found on perifollicular, small and large venule EC from GD and HT glands, that correlated with the severity of mononuclear infiltration. In addition, CD31 expression was observed in some intrathyroidal macrophages and T cells in close proximity to CD31+ EC. Furthermore, a markedly enhanced expression of endoglin, a transforming growth factor-beta binding protein, was mainly located on perifollicular EC and EC from small venules as well as in adjacent macrophages from GD and HT thyroid glands. This enhanced expression of E- and P-selectins, CD31 and endoglin by thyroid EC in GD and HT may reflect their ability to regulate leucocyte trafficking and activation.     

Mast cell and lymphoreticular infiltrates in neurofibromas. Comparison with nerve sheath tumors

Cellular heterogeneity produced by non-Schwannian elements may distinguish neurofibromas from other Schwann cell neoplasma and contribute to a different tumor biology. The present study compared cell counts of mast cells, T and B lymphocytes, and macrophages in 32 neurofibromas with those in 27 schwannomas, 9 malignant nerve sheath tumors, and 17 traumatic neuromas. Immunohistochemical and histochemical analyses were performed on formalin-fixed, paraffin-embedded tissues using two monoclonal antibodies against B-lymphocyte epitopes (LN-1 and LN-2), one monoclonal antibody against T-lymphocyte epitopes (UCHL-1), one polyclonal antibody recognizing alpha 1-antichymotrypsin (ACT), a macrophage/histiocytemarker, and toluidine blue O stains. Neurofibromas contained relatively high concentrations of mast cells significantly greater than the concentrations in other neoplastic or reactive nerve sheath tumors. Most neurofibromas also displayed moderate concentrations of LN-2 immunoreactive cells, similar to the concentrations in traumatic neuromas and not statistically different from cell counts in other tumor types. Limited, variable LN-1 and UCHL-1 immunoreactive infiltrates were detected in neurofibromas and some peripheral schwannomas. Rare or moderate ACT immunoreactivity was detected in the majority of neurofibromas, in contrast with the absence, or rare appearance, of ACT immunostaining in cranial and peripheral nerve schwannomas and moderate numbers of immunoreactive cells in many malignant nerve sheath tumors. Mast cells are an important cellular marker of neurofibromas and may participate in the pathogenesis of these neoplasms.     

Cutting Edge: The Etiology of Autoimmune Thyroid Diseases

Significant progress has been made in our understanding of the mechanisms leading to autoimmune thyroid diseases (AITD). For the first time, we are beginning to unravel these mechanisms at the molecular level. AITD, including Graves’ disease (GD) and Hashimoto’s thyroiditis (HT), are common autoimmune diseases affecting the thyroid. They have a complex etiology that involves genetic and environmental influences. Seven genes have been shown to contribute to the etiology of AITD. The first AITD gene discovered, HLA-DR3, is associated with both GD and HT. More recently, this association was dissected at the molecular level when it was shown that substitution of the neutral amino acids Ala or Gln with arginine at position beta 74 in the HLA-DR peptide binding pocket is the specific sequence change causing AITD. Non-MHC genes that confer susceptibility to AITD can be classified into two groups: (1) immune-regulatory genes (e.g., CD40, CTLA-4, and PTPN22); (2) thyroid-specific genes—thyroglobulin and TSH receptor genes. These genes interact with environmental factors, such as infection, likely through epigenetic mechanisms to trigger disease. In this review, we summarize the latest findings on disease susceptibility and modulation by environmental factors.     

Analysis of lectin binding in benign and malignant thyroid nodules

The lectin binding properties of ten cases each of adenomatoid nodule, follicular adenoma, and papillary carcinoma and five cases of microinvasive follicular carcinoma were examined histochemically and compared with adjacent normal thyroid tissue. Wheat germ agglutinin, concanavalin A, Ulex europaeus agglutinin I, peanut agglutinin, soybean agglutinin, Dolichos biflorus agglutinin, Ricinus communis agglutinin, and Helix pomatia agglutinin were employed. All the lectins but Ulex europaeus agglutinin I, peanut agglutinin, and Helix pomatia agglutinin were bound to thyroid parenchymal cells, colloid and stromal cells, but none uniquely to thyroid parenchymal cells. Helix pomatia agglutinin binding was present in stromal cells but not in parenchymal cells. Ulex europaeus agglutinin I binding to parenchymal cells was weakly positive only in five cases of papillary carcinoma. The binding in adenomatoid and neoplastic cells and their colloid was stronger than in adjacent normal thyroid tissue in all cases examined. Wheat germ agglutinin and concanavalin A binding was most intense among the lectins examined. In papillary carcinoma, lectin binding was observed mostly in the apical cytoplasm of carcinoma cells, whereas a diffuse surface binding pattern was predominant in follicular adenoma and carcinoma, adenomatoid nodules and normal thyroid gland. No consistent differences in lectin binding were found between follicular adenoma and carcinoma, or between adenomatoid nodules and follicular neoplasia.     

Immunohistochemical study of the lymph-follicles and lymphocytes in the gastric mucosa]

Immunohistochemical stainings according to ABC method (UCHL-1,L-26,MT-1,MB-1,IgG,IgA,IgD,IgM, kappa, lambda, LN-1 and LN-2) for the lymphocytes in the germinal center, mantle zone and infiltrative lymphocytes in the gastric mucosa of 30 cases of chronic gastritis and 10 cases of reactive lymphoreticular hyperplasia (RLH) were performed. Lymphocytes in the germinal center and mantle zone consisted usually of B-cells positively stained by L-26 and MB-1. However, in the interstitially infiltrative cells,T-cells positively stained by UCHL-1 and MT-1 were not infrequently contained. Immunoglobulin stainings revealed marked positivity for IgG,IgA,IgD,IgM, kappa and lambda in the interstitial lymphocytes and plasma cells. As to the RLH, small number of T-cells scattered in the germinal center and surrounding area of lymph follicles, and large number of T-cells were found among the follicles, where B-cells were more infrequently found than in the interstitial area of propria mucosae. Confusion of enlarged germinal centers and monotonous proliferation of lymphocytes among the lymph follicles showing monotonous positivity for the stains of heavy and light chains in the cases of RLH were suggestive of malignant change.     

High potential to tumor necrosis factor alpha (TNF-alpha) production of thyroid infiltrating T lymphocytes in Hashimoto's thyroiditis: a peculiar feature of destructive thyroid autoimmunity

T lymphocytes present in thyroid infiltrates of 6 patients with Hashimoto's thyroiditis (HT) and of 4 patients with Graves' disease (GD) were analyzed at clonal level and their profiles of mitogen-induced lymphokine secretion were characterized. Production of interleukin-2 (IL-2), interleukin-4 (IL-4), interferon-gamma (IFN-gamma) was measured in culture supernatants of a total number of 332 T cell clones (TCC) from HT, of 269 TCC from GD infiltrates and of 266 control TCC derived from normal lymphoid tissues. No significant difference was found in the ability to produce IL-2 between TCC from HT or GD infiltrates and control TCC. The proportion of HT- or GD-derived TCC able to produce IL-4 was extremely low (4 and 5%, respectively) in comparison with controls (19%). In contrast, the proportion of interferon-gamma (IFN-gamma)-producing (IFN-P) TCC derived from either HT (87%) or GD (80%) infiltrates was much higher (p less than 0.0005) than that found in controls (59%). In addition, most of IFN-P TCC from either HT or GD usually released higher amounts (p less than 0.002) of IFN-gamma than did control clones. No significant difference was found between GD infiltrates and controls in the proportions of TCC able to secrete TNF-alpha (39% and 47%, respectively), whereas the proportion of TNF-alpha-producing (TNF-P) TCC derived from HT (78%) was significantly higher (p less than 0.0001). In addition, most of both CD8 and CD4 TCC from HT released higher amounts of TNF-alpha than did TNF-P clones from controls or GD. These data suggest that T cells present in autoimmune thyroid infiltrates share a number of functions, such as high production of IFN-gamma, but differ with regard to their ability to secrete TNF-alpha, which is peculiar of most T cells present in the thyroid of HT patients.