Antibody reactivities to skeletal muscle proteins in a patient with lambda light chain secreting multiple myeloma, generalised amyloidosis and rhabdomyolysis.

Rhabdomyolysis is a rare complication in haematological malignancies, and a diverse range of factors has been implicated in the etiology of the syndrome. In the present study we analysed muscle morphology and antibody reactivities to skeletal muscle proteins in a patient diagnosed with lambda (lambda) light chain-secreting multiple myeloma (MM) and amyloidosis, who developed a progressive rhabdomyolysis. The muscle tissue analysis showed focal amyloid depositions and a low degree of atrophy and inflammation. Antibody reactivities against muscle proteins of approximately 42, 51 and 66 kD, respectively, were present in the patient's serum. The antibody specificities were revealed by lambda light chain- or IgM-specific antibodies. The results indicate a possible etiologic link between antibody reactivities towards muscle proteins and muscle tissue disorder in a patient with the unique combination of rhabdomyolysis, amyloidosis and MM of the light chain type.

Interleukin-10 gene promoter polymorphisms in multiple myeloma.

Multiple myeloma (MM) is a B-cell malignancy characterized by an accumulation of malignant plasma cells in the bone marrow. It is unclear whether genetic background could have an etiological impact on MM or influence the course of the disease. Interleukin-10 (IL-10) has been implicated in the growth and differentiation of normal B cells, and has also been shown to enhance the proliferation of MM cells. To address the putative involvement of IL-10 genetic variation in MM, we analyzed previously defined loci for bi-allelic polymorphism at position -1082 and two microsatellite loci (IL10.G and IL10.R) in the IL-10 promoter region. Seventy-three patients with MM, 27 with monoclonal gammopathy of undetermined significance, and 109 ethnically matched individuals as controls were included in the study. Significantly increased frequencies of the IL10.G genotype 136/136 and the IL10.R genotype 112/114, in addition to a decreased frequency of the IL10.R genotype 114/116, were found among the MM patients. Increased production of IL-10 was detected in the supernatants of lipopolysaccharide-stimulated peripheral blood mononuclear cells from MM patients who were homozygotes (136/136) and heterozygotes (136/non-136) for the IL10.G allele 136, as compared with the other IL10.G genotype carriers (non-136/non-136). These results suggest that the genetic variation in the IL-10 promoter region may play a role in the development of MM.

Cytotoxic T-lymphocyte antigen-4 microsatellite polymorphism is associated with multiple myeloma.

Multiple myeloma (MM) is a B-lineage malignancy with unknown aetiology. It has been considered that predisposing genetic factors might be implicated in the disease. In this study, the microsatellite polymorphism in the exon 3 of the cytotoxic T-lymphocyte antigen-4 (CTLA-4) gene was analysed in patients with MM and monoclonal gammopathy of undetermined significance (MGUS), together with ethnically matched healthy controls. The results showed that frequencies of the genotype 86/86 and of the allele 86 were significantly decreased in MM and MGUS compared with matched healthy controls, indicating that the CTLA-4 microsatellite polymorphism might represent a susceptibility locus for MM and MGUS.

Interleukin 6, tumour necrosis factor alpha, interleukin 1beta and interleukin 1 receptor antagonist promoter or coding gene polymorphisms in multiple myeloma.

Proinflammatory cytokines such as interleukin 6 (IL-6), tumour necrosis factor alpha (TNF-alpha) and IL-1beta are considered to be involved in the pathogenesis of multiple myeloma (MM). In the present study, we examined a G/C polymorphism at position -174 in the promoter region of IL-6, a biallelic polymorphism at position -308 in the promoter region of TNF-alpha, the TaqI restriction fragment length polymorphism in exon 5 of IL-1beta and a variable number of identical tandem repeat polymorphisms in intron 2 of IL-1 receptor antagonist (IL-1Ra) genes. The alleles of these loci are known to influence the level of production of the cytokines and the IL-1Ra. Seventy-three patients with MM, 27 with monoclonal gammopathy of undetermined significance (MGUS) and 129 healthy individuals were included. No difference was found between patients and healthy controls or between MM and MGUS patients in the distributions of genotypes and frequencies of alleles of the IL-6 (-174), TNF-alpha (-308), IL-1beta TaqI and IL-1Ra gene polymorphisms. No associations between the polymorphisms at the loci under study and clinical factors such as age, sex, clinical stage at onset and M-protein type were observed. Our results indicate that the cytokine (IL-6, TNF-alpha and IL-1beta) and IL-Ra gene polymorphisms do not confer susceptibility to the development of MM.

V-region-specific alterations of serum IgM production in multiple myeloma of IgG class.

INTRODUCTION: Multiple myeloma is accompanied by decreased polyclonal serum immunoglobulin concentrations. This suppression might be due to non-specific effects on the polyclonal lymphocyte populations as previously suggested, or it could include specific variable region-dependent mechanisms. Thus, differentiation, survival and activation to Ig secretion of B-lineage cells are dependent on the expression and signalling through the variable Ig receptor. The present study addresses the question whether such variable region-specific alterations were present in the peripheral repertoire of IgM antibodies in patients with IgG-secreting MM. MATERIALS AND METHODS: IgM reactivity repertoires towards a large panel of antigens in extracts of homologous tissues (liver, brain, stomach and heart muscle) and bacteria (Bacillus macquarensis) were analysed in sera from 22 patients diagnosed with IgG1 MM. Healthy, matched volunteers served as control donors. A modified Western assay was used, and values obtained from area integration by image analyses were submitted to multiparametric statistics. RESULTS AND CONCLUSION: The results confirm previous observations on the depression of serum IgM concentrations in multiple myeloma, and demonstrate concentration-independent, patient- and V-region-specific alterations in the polyclonal reactivity repertoires. Since the scoring of IgM reactivities by this technique is independent of IgG and because the deviations of IgM reactivity are not coincident with reactivities of (myeloma) IgG in the individual sera, the results indicate that the immunological syndrome of MM includes significant V-region-specific alterations in the polyclonal repertoires of IgM antibodies.

[Idiopathic hypophysitis in elderly patients. Report of 2 cases and review of literature]. / Hipofisitis idiopática en pacientes de edad avanzada. Presentación de dos casos y revisión de la literatura.

INTRODUCTION: Hypophysis inflammatory tumors are a non frequent cause for hypopituitarism. The motive of outpatient visit is headache. It is more frequent in pregnancy and immediate post-labor women. The pathology shows a lymphocytic inflammatory infiltrated or granulomas with giant cells (these can be accompanied with an infectious or autoimmune systemic disease associated). CLINICAL CASES: We present two elderly female patients, without infectious or autoimmune pathology associated, with suggestive abnormalities of hypophysial adenoma by MRI. Both presented a suspecting clinical manifestation of intracranial expansive lesion (progressive intensive headaches, with partial compromise of the III left pair in the second one), noticing by laboratory a subclinical hormonal dysfunction. Surgery of tumoral exeresis was underwent in both patients, showing a granulomatous hypophysitis in one and a lymphocytic hypophysitis in the other. CONCLUSION: Hypophysis inflammatory tumors must be considered as a preoperative differential diagnosis of every hypophysial tumor without important hormonal dysfunction associated, mainly in women and what is more in pregnancy or lying-in women, taking into account that surgery of tumoral resection can worsen a subclinic hypophysial dysfunction.

[MMP-2 expression (type IV collagenase) in gastric cancer]. / Expresión de MMP-2 (colagenasa tipo IV) en el carcinoma gástrico.

The protein MMP-2 (type IV collagenase) belongs to the family of metalloproteinases. Its function is related to cellular matrix degradation including basement membrane type IV collagen. Its presence in the neoplastic cells might enhance its capacity for dissemination. To find out some of its clinico-pathological and immunohistochemical behavior, 98 adenocarcinomas of the stomach were immunohistochemically studied, in search for MMP-2 in neoplastic cells. The results showed a correlation between MMP-2 with parietal depth of infiltration (p = 0.03) and with metastases in regional lymph nodes (p = 0.05). On the other hand, no correlation was found with sex, gastric localization, size of the tumor, histological type or grade neither with expression of MIB-1, c-erbB-2 nor p53 proteins, recurrence nor 5 year survival or no recurrency.

Expresion de MMP-2 (colagenasa tipo IV) en el carcinoma gastrico / MMP-2 expression (type IV collagenase) in gastric cancer

La MMP-2 (colagenosa tipo IV) es una proteína que pertence a la familia de las metaloproteinas, cuya función está relacionada con la degradación de la matriz extracelular. Su capacidad para degradar el colágeno IV de las membranas basales podría transformala en un agente facilitador de la diseminación neoplásica. Para ver su relación con algunas características clínico-patológicas e inmunohistoquímicas del cáncer gástrico se estudiaron 98 casos de adenocarcinoma de estómago determinado por inmunohistoquímica la presencia de MMP-2 en las células neoplásicas. Los resultados mostraron que había correlación entre la presencia de MMP-2 con el nivel de invasión parietal del tumor (p=0.03) y con la presencia de metástasis en ganglios regionales (p=0.05). En cambio no hubo asociación entre la expresión de MMP-2 con la frequencia por sexo, la localización dentro del estómago, el tamaño tumoral, el tipo histológico, el grado histológico, ni la expresión de las proteínas MIB-1, bcl-2, c-erbB-2 y p53. Tampoco se relacionó con la presencia de recidiva de la enfermedad ni con la sobrevida a los 5 años. (AU)

Expresion de MMP-2 (colagenasa tipo IV) en el carcinoma gastrico / MMP-2 expression (type IV collagenase) in gastric cancer

La MMP-2 (colagenosa tipo IV) es una proteína que pertence a la familia de las metaloproteinas, cuya función está relacionada con la degradación de la matriz extracelular. Su capacidad para degradar el colágeno IV de las membranas basales podría transformala en un agente facilitador de la diseminación neoplásica. Para ver su relación con algunas características clínico-patológicas e inmunohistoquímicas del cáncer gástrico se estudiaron 98 casos de adenocarcinoma de estómago determinado por inmunohistoquímica la presencia de MMP-2 en las células neoplásicas. Los resultados mostraron que había correlación entre la presencia de MMP-2 con el nivel de invasión parietal del tumor (p=0.03) y con la presencia de metástasis en ganglios regionales (p=0.05). En cambio no hubo asociación entre la expresión de MMP-2 con la frequencia por sexo, la localización dentro del estómago, el tamaño tumoral, el tipo histológico, el grado histológico, ni la expresión de las proteínas MIB-1, bcl-2, c-erbB-2 y p53. Tampoco se relacionó con la presencia de recidiva de la enfermedad ni con la sobrevida a los 5 años.

The repertoire of serum IgM in normal mice is largely independent of external antigenic contact.

Antigen-free (AGF) and germ-free (GF) mice, although essentially free of serum IgG, maintain normal levels of circulating IgM. Using a quantitative immunoblot assay, we have now analyzed the repertoire of serum IgM from AGF, GF, and specific pathogen-free (SPF) BALB/c mice, on large panels of natural antigens from homologous tissues and bacteria. The reactivity profiles were very similar in the three groups of mice. Multiparametric statistic evaluation of the data showed that BALB/c animals, SPF, GF, and AGF mice constitute an homogeneous group with similar immunoreactivity profiles when compared to C57BL/6. Differences between immunoreactivity profiles of GF and AGF mice were observed, but were not statistically significant. These results suggest that the serum IgM repertoire of normal mice is strictly regulated and selected by endogenous ligands.

Cytohistologic correlation of nuclear grade in breast carcinoma.

OBJECTIVE: To compare the nuclear grade (NG) in cytologic material (CNG) obtained from breast fine needle aspiration biopsies (FNABs) with the NG observed in surgical biopsies (BNG) of the same tumors. STUDY DESIGN: The study group consisted of 135 breast carcinomas with both FNAB and biopsy. Most of them were invasive ductal carcinomas. Cytologic aspirates and tissue sections were graded simultaneously by the three authors using a multiheaded microscope. Fisher's modification of Black's nuclear grading scheme was used. RESULTS: There was agreement between CNG and BNG in 70.37% of tumors. The percentage coincidence was slightly greater for NG 3. CONCLUSION: Nuclear grade can be easily established on FNAB. The lack of correlation (29.63%) may have been due to tumor heterogeneity and observer subjectivity when assigning nuclear grade.

Characteristic generated alterations of autoantibody patterns in idiopathic thrombocytopenic purpura.

Using a Western blot technique that allows quantitative detection of antibody reactivities to a large number of antigens, serum IgG and IgM antibody repertoires were compared in a group of 19 patients with a diagnosis of idiopathic thrombocytopenic purpura (ITP) and respective healthy controls. The results show that, irrespective of the duration of thrombocytopenia, age of the patients, and type of therapy, all ITP donors share characteristic alterations of serum antibody reactivity patterns on homologous erythrocyte and liver antigens. Multiparametric analyses of the immunoreactivity data readily segregated the groups of ITP and healthy donors. Similar analyses also distinguished ITP sera from those of a group of patients with systemic lupus erythematosus (SLE). We conclude that ITP is an autoimmune disease associated with generalized alterations of antibody repertoires, that may be characteristic enough to allow for diagnosis.

Instability of natural antibody repertoires in systemic lupus erythematosus patients, revealed by multiparametric analysis of serum antibody reactivities.

Recent views on autoimmune diseases invoke generalized but specific perturbations in antibody repertoires, rather than the clonally restricted or non-specific polyclonal alterations proposed thus far. The present experiments analyse serum antibody reactivities in 24 systemic lupus erythematosus (SLE) patients and 17 healthy controls, using a method that quantitatively scores a large number of antibody reactivities and allows for multiparametric statistical analyses. The results show global but relatively specific perturbations in SLE antibody repertoires, and identify novel disease-associated reactivity patterns. Furthermore, a time series analysis of serum antibodies over 3 months demonstrates instability of natural antibody repertoires in individual SLE patients, contrasting with their remarkable conservation in healthy donors. Moreover, the method used clusters controls and patients independently, and might prove of diagnostic value, once large data bases are established.

[Expression of p53 and bcl-2 in colorectal adenomas and carcinomas]. / Expresión de p53 y de bcl-2 en adenomas y carcinomas colorrectales.

Recent publications have associated p53 and bcl-2 genes in the process of neoplastic transformation. As the colonic adenoma-carcinoma sequence is an adequate natural model for carcinogenesis, it was considered interesting to analyze the expression of bcl-2 and p53 in these neoplasms. Seventy three adenomatous polyps (adenomas) and 60 adenocarcinomas of the colon and rectum were studied. Adenomas showed mild dysplasia in 16, moderate in 27, severe in 15 and focal carcinoma in the remaining 15. Adenocarcinomas surpassed the deep muscle layer in every case and were moderately differentiated. The studied gene expression was analized immunohistochemically using antibodies bcl-2 from Dako and p53 from Novocastra, both at a 1:100 dilution. Cytoplasmic stain for bcl-2 and nuclear stain for p53 above 10% of the cells were considered positive for each gene respectively. Results showed that there was accumulation of p53 protein in 26/58 (45%) adenomas with different grades of dysplasia. This result is similar to the reactivity found in adenomas with focal carcinoma where 8/15 (53%, p = 0.4) were positive but different from adenocarcinomas which were positive in 47/60 (78%, p = 0.0001). Regarding bcl-2, positivity was found in 53/73 (73%) of all the adenomas whereas adenocarcinoma showed expression in 14/60 (23%, p = 0.0000). When adenomas were grouped according to their degree of dysplasia and the existence of focal carcinoma, a diminishing frequency of reactivity for bcl-2 was found and when adenomas with three different grades of dysplasia were fused together, 47/58 (81%) were positive and this was compared with adenomas having focal carcinoma, 6/15 (40%) and with adenocarcinoma, 14/60 (23%), they showed significant differences (p = 0.001 and p = 0.0000 respectively). The analysis of the frequency of expression for both genes studied in the different lesions described yielded an inverse relation between them. This study allows the conclusion that the expression of bcl-2 is an early event in carcinogenesis and that it is replaced by mutation of p53 as the neoplastic change progresses.

Carcinoembryonic antigen expression in stages I and II breast cancer: its relationship with clinicopathologic factors.

Carcinoembryonic antigen (CEA) has been detected by immunohistochemistry in breast carcinoma, but its relationship with prognosis is still unclear. This difficulty may be because of the great variety of antibodies used for its determination. In the present study, 271 stages I and II breast carcinomas are analyzed by immunohistochemistry, using T84.66 antibody, a well-known highly specific CEA antibody. The results show that CEA expression was not associated with any of the clinicopathologic factors analyzed. Factors associated with disease-free survival (DFS) after univariate logistic regression analyses were tumor size smaller than 2 cm (P = .01), lymph node free of metastases (P = .0000), low nuclear grade (P = .007), absence of c-erbB-2 overexpression (P = .02), and bcl-2 (P = .005) and CEA expression (P = .005), whereas those significantly associated with a better overall survival (OS) were tumor size small than 2 cm (P = .002), lymph node free of metastases (P = .0001), low nuclear grade (P = .01), low histological grade (P = .02), absence of c-erbB-2 overexpression (P = .002) and bcl-2 expression (P = .01). After multivariate stepwise regression analysis, lymph node free of metastases (P = .0000), CEA expression (P = .001), absence of c-erbB-2 overexpression (P = .01), and bcl-2 expression (P = .01) were found to be independent factors associated with DFS, whereas lymph node free of metastases (P = .0000), tumor size smaller than 2 cm (P = .0000), and absence of c-erbB-2 overexpression (P = .004) were associated with a better OS. These results show that immunohistochemical detection of CEA with the antibody T84.66 may be useful as an additional factor in establishing breast cancer prognosis.

B-lineage cell deficits in bone marrow of lpr/lpr mice.

Analyses of bone marrow (BM) lymphocytes in C57BL/6 mice homozygous for the lpr mutation (B6.lpr) disclosed low numbers of pre-B and B cells, as compared with age-matched control B6 mice. BM depletion in B6.lpr mice was selective for B-lineage cells, appeared in young adults, and developed markedly with age and disease progression, contrasting with the peripheral lymphocyte hypercellularity. Normalization of pre-B and B cellularity in BM of B6.lpr mice was observed after administration of polyclonal Ig, that also markedly improved the clinical condition. Isolated pre-B (B220+ IgM-) cells from B6 or B6.lpr mice, however, showed essentially the same rates of IL-7-dependent proliferation and differentiation to B (IgM+) cells in culture, indicating that the BM B-lineage deficit is not the result of an intrinsic defect in B cell generation.

[Reproducibility of nuclear grade in breast neoplasm. A multicenter experience]. / Reproducibilidad del grado nuclear en el cáncer de mama. Una experiencia multicéntrica.

Nuclear grade is considered a valuable prognostic factor in mammary carcinomas. Since the histological diagnosis of most of these tumors is made by "non expert" pathologists, it was considered interesting to find out the reproducibility of general pathologists to define the nuclear grade. In order to do this, a series of 15 mammary carcinomas, 10 of them randomly selected and 5 because they were considered difficult to classify for nuclear grade, were examined separately by 10 general pathologists. In a first round of observation, each one of them graded the cases according to their own criteria as used routinely, and for a second round they followed a written guide. An analysis of variance was applied to the data and no significant differences were found between observers, neither in the randomly selected cases nor in the total series. The written guide, surprisingly, instead of lowering the differences, increased them. Analysis of the individual performance of observers showed two of them having a great variation between both rounds of observation, and this was considered to influence the results of the whole group. Interobserver performance to discriminate high grade tumors (G3) from the rest, showed a good correlation in all the participants. These results allow us to conclude that in this series, examined by general pathologists, an acceptable reproducibility was observed, specially when high risk tumors were being identified.

[Immunohistochemical characteristics of mammary carcinomas with estrogen-negative and progesterone-positive receptors]. / Características de los carcinomas de mama receptores de estrógenos negativos y receptores de progesterona positivos determinados por inmunohistoquímica.

In a series of 256 mammary carcinomas, 22 (8.5%) were positive for progesterone receptors (PR) and negative for estrogen receptors (ER). These cases seem to belong to a distinctive group with a biologic behavior not well understood. In order to contribute to a better understanding of such tumors, their association with different pathologic and immunohistochemical factors were compared with those of the rest of the tumors of the series. The results were that favorable factors such as smaller size, negative axillary lymph nodes and low histologic and nuclear grades were decreasingly associated with tumors that were ER+ PR+; ER+ PR-; ER- PR+; and ER- PR-. In relation to immunohistochemical features, tumors that were ER+ PR+; ER+ PR- and ER- PR+ behaved in a similar way, whereas ER- PR- tumors were different from the rest because fewer expressed bcl-2 (p = 0.0000) and had a greater expression for p53 (p = 0.009) and MIB-1/Ki-67 (p = 0.05). No significant differences were found between the four populations in recurrence rate or metastases, nor overall survival. In conclusion, these findings show that tumors that are ER- PR+ might have biological characteristics somewhere in between ER+ PR+ and ER- PR+.

Diagnostic value of p53 protein in the study of serous effusions.

The differential diagnosis between neoplastic and reactive mesothelial cells is one of the most frequent problems in the study of serous effusions. We assessed the utility of the immunohistochemical determination of p53 protein as a marker of malignancy in 34 embedded blocks of neoplastic fluids and 30 nonneoplastic effusions. Eleven (32.4%) of the tumor fluids were positive for this antibody, while all the nonneoplastic fluids were negative. A specificity of 100% and a sensitivity of 59% were observed. The immunohistochemical determination of p53 protein seems to be helpful in the differential diagnosis of effusions; its principal limitation is its relatively low sensitivity.