Germinal centers and the B-cell system

Summary The migration pattern of germinal center cells of the rabbit appendix was studied and compared with that of appendix dome cells, spleen cells, thymus cells and thoracic duct lymphocytes. To discriminate T-and B-cell migration pathways, normal or T-cell-depleted rabbits were used as donors. Cell suspensions were labeled in vitro with ³H-leucine followed by intravenous transfer. The migration of labeled cells in lymphoid organs was studied using autoradiography, particular attention being paid to the spleen of the recipient. B-cells from the appendix dome, spleen and thoracic-duct lymph migrate to primary follicles or the corona of secondary follicles via thymus-dependent areas of peripheral lymphoid organs. In contrast, a B-cell subpopulation from the germinal centers of the appendix migrates to the center of splenic primary follicles and into germinal centers. The migration of germinal center cells to splenic follicle centers is not enhanced by specific antigens. The migration properties of B-cells, possibly changing during differentiation, may be instrumental in the two types of immune reactions, i.e., plasma-cell reaction and germinal-center reaction.     

Age-related changes in the morphology and the distribution of IgA and IgG in the pharyngeal tonsils of yaks (Bos grunniens)

To evaluate age-related changes in the morphology as well as the expression and localization of IgA and IgG in yak pharyngeal tonsils, 20 healthy yaks were divided into four age groups [newborn (1–7 days old), juvenile (5–7 months old), adult (3–6 years old) and old (7–10 years old)]. Morphologic characteristics were observed by histological techniques. The expression and localization of IgA and IgG in pharyngeal tonsils were detected by enzyme linked immunosorbent assay (ELISA) and immunohistochemistry, respectively. The results showed that the epithelium of the pharyngeal tonsils included nonreticular epithelium with an intact basement membrane and reticular epithelium with a discontinuous basement membrane and nonepithelial cell infiltration. In newborn yaks, only primary lymphoid follicles were observed in pharyngeal tonsils. In other age groups, both primary and secondary lymphoid follicles were observed, but some of the lymphoid follicles in the old yaks were degenerated. The number of lymphoid follicles increased from the newborn to the adult group and peaked in the adult group, but the number decreased in the old group. In addition, the age-related trends of IgA and IgG protein expression were similar to those of the number of lymphoid follicles. The concentration of IgG was significantly higher than that of IgA in all age groups. Both IgA and IgG antibody secreting cells (ASCs) were distributed in the subepithelial region of the nonreticular epithelium, the reticular epithelium, the lymphoid follicles, the interfollicular areas and in between the salivary glands. The densities of IgA and IgG ASCs in pharyngeal tonsils were similar to the expression trend of both proteins in each age group. The results indicate that the morphology and amount of lymphoid follicles in yak pharyngeal tonsils vary with age. Pharyngeal tonsils produce more IgG than IgA, indicating that IgG could be significant component of mucosal immune responses in yaks.     

SALIVARY GLAND TUMORS

In a review of a series of 248 salivary gland tumors, seen over a 28-year period, all pathologic material was brought up to date by reclassification according to more recent criteria and nomenclature. In parotid tumors, a probable lowered recurrence rate and a definite decrease in incidence of permanent facial nerve paralysis was found in the more recent cases in which the "Y" incision was used, with identification of the seventh nerve as it leaves the stylomastoid foramen. The five-year recurrence rate for primary mixed tumor was 8.3 per cent, and in recurrent cases it was found to be 18.1 per cent. Of 44 patients with malignant salivary gland tumors in all sites who were observed for five years or more, 32 or 72.7 per cent survived five years.     

A comparison of euglobulin fractions and whole serum in the hemagglutination and latex fixation tests

One hundred and thirty-two sera were investigated with the Waaler-Rose and latex fixation reactions. The reactions were performed with serum, with acid-precipitated euglobulin, and with cold-precipitated euglobulin. The material consisted of 35 sera from healthy persons, 23 from patients with various diseases, 28 from patients with joint symptoms not due to rheumatoid arthritis, and 46 from patients with classical rheumatoid arthritis.In rheumatoid arthritis sera, an increase in positive reactions was obtained in the Waaler-Rose test from 70 per cent in serum to 83 per cent in acid-precipitated euglobulin. This increase was due to a greater specificity of reactions with low titers. The cold-precipitated euglobulin gave less positive Waaler-Rose reactions than the acid-precipitated euglobulin. With the latex fixation test an increase from 65 per cent positive reactions in serum to about 71 per cent with both cold- and acid-precipitated euglobulin fractions was obtained. Here, the increase consisted of reactions negative in serum but positive in the euglobulin fractions, but again with low titers. Because the increase in positive reactions consists merely of low titer values, fractionation of sera only slightly enhances the reliability of the serological tests.Negatively reacting rheumatoid arthritis sera often had low values of the _2A globulin.     

IFN-γ plays an essential role in the pathogenesis of gastric lymphoid follicles formation caused by Helicobacter suis infection

In this study, we aimed to assess the role of helper T cells in the development of gastric lymphoid follicles induced by Helicobacter suis infection. C57BL/6J mice were orally inoculated with H. suis. Six weeks after infection, gastric lymphoid follicles were observed in the gastric mucosa by hematoxylin and eosin staining, and the number of follicles was increased throughout the infection period. An immunohistological examination showed that the lymphoid follicles were composed of B cells, CD4-positive helper T cells, and dendritic cells (DC). It was also revealed that the mRNA expression level of interferon-γ (IFN-γ) in the gastric mucosa was significantly increased at 12 weeks after infection. No gastric lymphoid follicles were detected in IFN-γ-deficient mice that had been infected with H. suis at 12 weeks after infection, although the development of lymphoid follicles in IL-4-deficient mice infected with H. suis was similar to that seen in the wild-type mice. In conclusion, IFN-γ, a Th1 cytokine, is deeply involved in the pathogenesis of gastric lymphoid follicles induced by H. suis infection, and it is suggested that CD4-positive T cells and DC aid in the expansion of gastric lymphoid follicles.     

Ultrastructural and cytoarchitectural features of lymphoreticular organs in the colon and rectum of adult BALB/c mice

The structure and function of colonic mucosal lymphoid organs remain largely unexplored, especially in the rectum hidden within the pelvic vault. Two-month-old female BALB/c mice were anesthetized, and the entire colon was removed from cecum to anus. Distal colonic patches were then prepared for electron microscopy or were quick-frozen and sectioned for immunoperoxidase localization of B cells and T cell subsets. Aggregated lymphoid follicles were distributed irregularly along the entire colon with an average of 1.4 patches per centimeter of colon length. There were large collections of follicles opposite the ileocecal valve (cecal patches), variable numbers of patches throughout the colon, and at least one patch within 10 mm of the anus (rectal patch). Follicles were adjacent to branching crypts lined by epithelium infiltrated by lymphoid cells and containing few goblet cells. In electron micrographs, M cells were identified by their short, irregular microvilli; intraepithelial lymphoid cells; reduced lysosomal dense bodies; and an expanded tubulovesicular network. Small germinal centers were seen. Cytoarchitectural components of colonic lymphoid follicles and Peyer's patch follicles were remarkably similar, despite differences in surrounding mucosa and luminal microbial exposure. The presence of organized lymphoid tissue with M cells and germinal centers suggests that transepithelial particle transport and antigen recognition can take place in the rectum. Whether such tissue has the capacity for uptake of luminal microorganisms is of particular interest, not only because colonic follicles may be sites for local initiation of immune responses but also because they may be important entry points for systemic infection.     

Fine needle aspiration cytology of bone lesions. Analysis of a three-year experience in rural Africa

FNA biopsies were performed on 24 patients with bone lesions. Cytology diagnosed ten of the cases as classic Burkitt's lymphomas. The cytodiagnoses in the remaining 14 cases were primary bone tumors (5 cases), bone cysts (2 cases), inflammatory lesions (4 cases) and inadequate material (3 cases). Smears of the Burkitt's lymphomas of the jaw contained starry sky macrophages and neoplastic lymphoid cells with deep basophilic cytoplasms and fine vacuolizations. Taking into consideration the load of Burkitt's lymphoma cases in Africa, FNA cytology appears to be a very simple method for getting a quick tissue diagnosis (results were available within 24 hours). For doctors working in tropical hospitals with limited facilities, FNA cytopathology is very useful for distinguishing between tumors and inflammations and for differentiating between benign and malignant tumors.     

Results of irradiation in cancer of the lip,tongue and ear

CANCER OF THE LIP: The primary lesion can be controlled by irradiation in approximately 80 per cent of cases. For lesions with metastases there is only about a 25 per cent chance of five-year arrest (irradiation of the primary lesion followed by excision of involved nodes). CANCER OF THE TONGUE: Lesions in the anterior two-thirds are controllable by irradiation in about 50 per cent of cases if the nodes are not involved; the salvage is only about 15 per cent if the nodes are involved (nodes treated surgically). Lesions in the posterior third of the tongue are seldom controlled in the author's experience. CANCER OF THE EAR (AURICLE): Five-year arrest of basal-cell lesions should be attained by irradiation in about 80 per cent of cases; of squamous-cell lesions in about 60 per cent. If the lesion is extensive, radiation does not offer a superior cosmetic result to operation and entails danger of late chondronecrosis. Therefore extensive lesions are probably best treated surgically. In either event, it appears probable that results of irradiation can be improved by the use of more adequate fields and greater fractionation.     

Damage to secondary lymphoid follicles in AIDS-related persistent generalized lymphadenopathy, as revealed by the behaviour of dendritic reticulum cells possessing immunoreactive acid cysteine-proteinase inhibitor

It has been shown previously that dendritic reticulum cells (DRC) in human secondary lymphoid follicles possess an immunoreactive acid cysteine-proteinase inhibitor (ACPI). In the present study, lymph nodes from 12 patients with AIDS-related persistent generalized lymphadenopathy (PGL) were investigated in order to detect whether or not any alterations occur in ACPI-immunoreactive DRC in this disorder. In the majority of PGL cases, profound alterations were found, the main characteristics of which were erosion, partial or total disruption of lymphoid follicles. However, similar though much less marked alterations were also found in some control cases. It is concluded that this type of follicular damage is a common and characteristic feature in PGL. It is not specific to PGL, however, but represents rather a special type of reaction in lymphatic tissue. The advantage of ACPI immunohistology for demonstrating the DRC pattern is that it can be performed on routinely fixed and paraffin-embedded tissues.     

Prostatectomy in a county hospital; a review of 677 cases in a six-year period

Transurethral procedures were used in 620 of 677 cases in which prostatectomy was done (principally by residents supervised by a urologist) at a county hospital in a six-year period. Open operations were used in the other 57 cases. Results were classified as "excellent" in 46.8 per cent of the transurethral cases and as "good" in 36.3 per cent.     

Classification of small type B/C follicles as primordial follicles in mature rats

In the present study, follicles were classified according to the morphology of their granulosa cells. Type B follicles contained only flattened granulosa cells; type B/C follicles had a mixture of flattened and cuboidal granulosa cells in a single layer, and type C follicles had a single layer of cuboidal granulosa cells. The primary objectives of the study were to determine whether 5-bromo-2-deoxyuridine incorporation into type B/C follicles was a marker for initiation of growth and how long type B/C follicles could remain at the same stage before transformation to type C follicles. Female Holtzman rats received bromo-deoxyuridine for 7 days. After the infusion (day minipumps were removed = day 0), rats were ovariectomized on days 0 (n = 9), 30 (n = 8), 90 (n = 8) and 150 (n = 9). The numbers of type B, B/C and C follicles within one ovary were determined using modified fractionator counting. Analysis over all times demonstrated that there were more (P < 0.0001) type B/C (941 +/- 61 per ovary) than type C (140 +/- 18 per ovary) or type B (159 +/- 19 per ovary) follicles. The numbers of type B and type C follicles did not differ from each other at any time. Only one of 34 rats evaluated had bromo-deoxyuridine-labelled type B follicles. On day 150, 57% of the bromo-deoxyuridine-labelled type B/C follicles remained from day 0. It is concluded that (1) DNA synthesis in granulosa cells of type B/C follicles is not a reliable indicator of impending growth; and (2) type B and type B/C follicles are both components of the pool of primordial follicles.     

INFLUENZA VACCINE,ASIAN STRAIN—Reactions Following Its Use in Adults

A study of reactions following influenza vaccine inoculation of 327 employees of Peralta Hospital, 55 men and 272 women, showed a very low value for significant or severe reactions. The reaction rate as observed with the present monovalent vaccine containing 200 CCA units of Asian strain, Type A influenza virus, was considerably lower than that reported with previous polyvalent vaccines containing up to 1,400 or 1,500 CCA units of total virus content.The absenteeism rate was 1.1 per cent for women, nil for men.The incidence of reactions was much greater in women than in men. Local reactions such as pain, swelling, or redness at the site of injection occurred in 29.1 per cent of men and 35.7 per cent of women. The incidence of systemic reactions—fever, aching, chilliness, headache, nausea and vomiting—was 3.6 per cent in men and 8.8 per cent in women. About 9 per cent of men and 30 per cent of women had both local and systemic reaction. Some 58 per cent of men and 25 per cent of women had no reaction.The greater majority of reactions appeared within five hours after inoculation with influenza vaccine.In adults the prevention of anaphylactic reactions due to the small amount of egg protein in influenza vaccine, can be accomplished by screening for history of hypersensitivity to egg, chicken or chicken feather. In questionable cases, intradermal testing can be done.The reaction rate observed in this study for the present influenza vaccine was so low that it ought not deter immunization.     

Revision of a nonunited subtrochanteric femoral fracture around a failed intramedullary nail with the use of RIA products, BMP-7 and hydroxyapatite: a case report

Introduction

Küttner's tumor is characterized through histology by peri-ductal fibrosis, dense lymphocytic infiltration with lymphoid follicles, loss of acini, and occasional marked sclerosis of the salivary gland. On occasion, Küttner's tumor can be difficult to distinguish from malignant neoplasm.

Case presentation

A 58-year-old Japanese man was referred to our hospital with a three-month history of a painless swollen mass in the right sub-mandibular region. Histological findings revealed both lymphoid follicles with reactive germinal centers and variously sized lymphoid follicle-like nodules without definitive germinal centers or mantle zones. B-cells of similar size and shape occupied the lymphoid follicle-like nodules and stained positive for B-cell lymphoma. These cells were detected in the polyclonal B-cells by flow cytometric analysis and tested negative for CD10. Unusual B-cell proliferation was observed, but as there was no definitive evidence of B-cell lymphoma, the lesion was diagnosed as Küttner's tumor.

Conclusion

We report on a rare case of Küttner's tumor associated with fibrosclerosis and atypical lymphoid hyperplasia in both the sub-mandibular gland and regional lymph nodes. Although more cases need to be investigated, our findings might be helpful to further studies seeking to clarify the etiology of idiopathic sclerosing lesions arising in the organs and regional lymph nodes.     

Cigarette smoke-induced pulmonary emphysema in scid-mice. Is the acquired immune system required?

Background

Chronic obstructive pulmonary disease is associated with a chronic inflammatory response of the host to chronic exposure to inhaled toxic gases and particles. Although inflammatory cells of both the innate and adaptive immune system infiltrate the lungs in pulmonary emphysema and form lymphoid follicles around the small airways, the exact role of the acquired immune system in the pathogenesis of emphysema is not known.

Methods

In this study, wild type Balb/c mice and immunodeficient scid mice – which lack functional B- and T-cells – were exposed to mainstream cigarette smoke (CS) for 5 weeks or 6 months.

Results

Subacute CS-exposure for 5 weeks significantly increased innate inflammatory cells (neutrophils, macrophages and dendritic cells) in the bronchoalveolar lavage (BAL) fluid of wild type mice and scid mice, which correlated with the CS-induced upregulation of the chemokines Monocyte Chemotactic Protein-1, Macrophage Inflammatory Protein-3α and KC (= mouse Interleukin-8). Chronic CS-exposure for 6 months significantly increased the number of neutrophils, macrophages, dendritic cells, CD4⁺ and CD8⁺ T-lymphocytes in BAL fluid and lungs of wild type mice compared to air-exposed littermates, and augmented the size and number of peribronchial lymphoid follicles. In contrast, neither B-lymphocytes, nor T-lymphocytes, nor lymphoid follicles could be discerned in the lungs of air- or CS-exposed scid mice. Importantly, chronic CS-exposure induced pulmonary emphysema in both wild type animals and scid mice, as evidenced by a significant increase in the mean linear intercept and the destructive index of CS-exposed versus air-exposed animals. The CS-induced emphysema was associated with increased mRNA expression of matrix metalloproteinase-12 in the lungs and increased protein levels of Tumor Necrosis Factor-α in the BAL fluid of CS-exposed Balb/c and scid mice compared to air-exposed littermates.

Conclusion

This study suggests that the adaptive immune system is not required per se to develop pulmonary emphysema in response to chronic CS-exposure, since emphysema can be induced in scid mice, which lack lymphoid follicles as well as functional B- and T-cells.     

Pathology of supraclavicular lymphadenopathy in Chandigarh, north India: an audit of 200 cases diagnosed by needle aspiration

OBJECTIVE: Fine needle aspiration cytology (FNAC) of lymph nodes can be used routinely as a first-line diagnostic test. The majority of studies reveal a malignant cause for palpable supraclavicular lymph nodes. The present audit further emphasizes the use of FNAC as a first-line investigation for the evaluation of enlarged supraclavicular lymph nodes. METHODS: A total of 200 cases of palpable supraclavicular lymph node(s) were included in the present study. RESULTS: Left supraclavicular lymph nodes were found to be more commonly involved (59.5% cases). Sixty-four per cent cases showed metastatic deposits and 13.5% cases were diagnosed as tuberculosis. Ten per cent cases showed reactive lymphoid hyperplasia; 0.5% (one) case showed only necrosis and on autopsy, a microscopic focus of choriocarcinoma was found in the testis. In 7.5% cases, diagnostic material could not be aspirated despite repeated attempts. Common metastatic tumours were from lung (22% cases), breast (16.4% cases), cervix (11% cases) and oesophagus (8.6% cases). In 13.3% cases the primary site was unknown and the diagnosis of malignancy first came from FNAC. CONCLUSION: The present study further highlights the importance of FNAC as a first-line diagnostic modality in the evaluation of supraclavicular lymphadenopathy. A full history, radiological findings and immunochemistry in difficult cases can help to arrive at a definitive diagnosis.     

Experience with using aclarubicin in the treatment of acute leukemia and blast crisis of chronic myeloid leukemia]

The clinical efficacy of aclarubicin, an anthracycline antibiotic, was studied in 48 patients with leukemia. The antibiotic was used in the following combinations with cytarabine: "7 + 7", "5 + 5" and "7 & 3". A complete remission was stated in 14 (42.4 per cent) out of 33 patients with acute nonlymphoid leukemia, 6 (43 per cent) out of the 14 patients having relapses. The combined therapy was effective in 4 out of 5 pre-resistant patients. The "7 + 3" scheme was the most beneficial. The most common adverse reactions were nausea and vomiting.     

CEA, ZGM and EMA localization in cells of pleural and peritoneal effusion: a preliminary study

Carcinoembryonic antigen (CEA), the zinc glycinate marker (ZGM) and epithelial membrane antigen (EMA) have been described as epithelial or tumour markers of varying specificity. These antigens were studied by immunoperoxidase localization in selected cell blocks of 62 pleural or peritoneal effusions and compared to cytological findings and review of the clinical records. By cytological criteria, 25 of the cell blocks were positive for malignancy, 30 negative, and 7 inconclusive. CEA, ZGM, and EMA by immunoperoxidase staining were localized on the cell surface and often in the cytoplasm of malignant cells, in 11/25 (44 per cent), 17/25 (68 per cent) and 22/25 (88 per cent) of the positive cell blocks respectively. Ten (40 per cent) of these cases were positive for all three antigens, 7 (28 per cent) for two, and 6 (24 per cent) for one. Of the 7 cases which were inconclusive on routine cytological reporting, 5 were positive for at least one marker. In 3 of the 5 a diagnosis of malignancy was confirmed, and in the other two was strongly suspected as malignant on clinical grounds. Macrophages were sometimes positive for one or more markers (but showed cytoplasmic staining only) and mesothelial cells in some cases stained positively for EMA but were always negative for CEA and ZGM. Localization of the 3 antigens in cells of malignant effusions was compared with their localization in the primary tumours in 9 cases. Localization corresponded for CEA in 7 of 9 cases, for EMA in 8 of 8 an for ZGM in only 2 of 9. Effusion fluid levels for CEA were compared with the cytological and immunocytochemical findings in 30 cases. Mucin stains performed on the cell blocks were also compared with the immunoperoxidase findings.     

RESULTS OF IRRADIATION IN CANCER OF THE LIP,TONGUE AND EAR

Cancer of the lip: The primary lesion can be controlled by irradiation in approximately 80 per cent of cases. For lesions with metastases there is only about a 25 per cent chance of five-year arrest (irradiation of the primary lesion followed by excision of involved nodes).Cancer of the tongue: Lesions in the anterior two-thirds are controllable by irradiation in about 50 per cent of cases if the nodes are not involved; the salvage is only about 15 per cent if the nodes are involved (nodes treated surgically). Lesions in the posterior third of the tongue are seldom controlled in the author''s experience.Cancer of the ear (auricle): Five-year arrest of basal-cell lesions should be attained by irradiation in about 80 per cent of cases; of squamous-cell lesions in about 60 per cent. If the lesion is extensive, radiation does not offer a superior cosmetic result to operation and entails danger of late chondronecrosis. Therefore extensive lesions are probably best treated surgically. In either event, it appears probable that results of irradiation can be improved by the use of more adequate fields and greater fractionation.     

B lymphocyte autoimmunity in rheumatoid synovitis is independent of ectopic lymphoid neogenesis

B lymphocyte autoimmunity plays a crucial role in the pathogenesis of rheumatoid arthritis. The local production of autoantibodies and the presence of ectopic lymphoid neogenesis in the rheumatoid synovium suggest that these dedicated microenvironments resembling canonical lymphoid follicles may regulate the initiation and maturation of B cell autoimmunity. In this study, we assessed experimentally the relevance of ectopic lymphoid neogenesis for B cell autoimmunity by a detailed structural, molecular, and serological analysis of seropositive and seronegative human synovitis. We demonstrate that synovial lymphoid neogenesis is a reversible process associated with inflammation which is neither restricted to nor preferentially associated with autoantibody positive rheumatic conditions. Despite the abundant expression of key chemokines and cytokines required for full differentiation toward germinal center reactions, synovial lymphoid neogenesis in rheumatoid arthritis only occasionally progresses toward fully differentiated follicles. In agreement with that observation, we could not detect Ag-driven clonal expansion and affinity maturation of B lymphocytes. Furthermore, ectopic lymphoid neogenesis is not directly associated with local production of anti-citrullinated protein Abs and rheumatoid factor in the rheumatoid joint. Therefore, we conclude that synovial lymphoid neogenesis is not a major determinant of these rheumatoid arthritis-specific autoantibody responses.     

Migration of B lymphocytes in lymphoid organs of lethally irradiated,thymocyte-reconstituted mice

Summary The migration of radiolabeled intravenously injected B lymphocytes through thymus-dependent areas was studied in lymphoid organs of mice with experimentally defined T cell domains (B cell-deprived mice or T mice). In the spleen, B cells were found to enter the peri-arteriolar lymphoid sheath (PALS) by two routes: (i) via the marginal zone, and (ii) via reticulin sheaths surrounding terminal arterioles. B cells migrated through the peripheral and central PALS and initiated the formation of primary follicles in the peripheral PALS 6 h after injection. Distinct primary follicles were noted at 18 h after injection of the labeled B cells. After 24 h small numbers of labeled cells were also noted in the efferent lymphatic vessels of the spleen.The reconstitution of B cell compartments in the mesenteric lymph node was delayed compared to the spleen. B cells entered the nodal stroma across the wall of high endothelial venules in the paracortex and by 6 h were found scattered throughout the paracortex. Isolated clusters of a few labeled cells were noted in the outer cortex at 18 h after cell transfer. Defined primary nodules were seen only 24 h after reconstitution. A minority of labeled cells was found at 24 h in the cortico-medullary junctions and in medullary cords.The present study shows that B lymphocytes traverse T cell domains on their way to their own specific B cell compartments. The immunological significance of this particular migration route is discussed in view of data on the cellular cooperation of B cells, T cells and macrophages during the humoral immune response.