Respuesta anticlases mayores de anticuerpos humanos séricos en antisueros hiperinmunes obtenidos en carneros / Characterization of hyperimmune anti-human serum produced in sheep

Introducción: La inmunoelectroforesis es una técnica de precipitación que permite la caracterización de muestras biológicas complejas. En el Departamento de Inmunología del Instituto de Ciencias Básicas y Preclínicas Victoria de Girón se cuenta con un antisuero hiperinmune obtenido por inmunizaciones de carneros contra proteínas totales séricas humanas y con otro antisuero anti IgA de calostro humano. Objetivo: Identificar IgG, IgM e IgA en suero humano y determinar respuesta anti IgM humana en el antisuero anti IgA de calostro humano obtenido en carnero. Material y Métodos: Se realizó un estudio observacional, descriptivo y transversal desde noviembre de 2017 hasta junio de 2018. Se desarrolló una inmunoelectroforesis de suero humano normal empleando el antisuero hiperinmune. Resultados: Se identificaron IgG, IgM e IgA además de albúmina y otras fracciones proteicas y se determinó respuesta anti IgM humana en el antisuero anti IgA de calostro humano obtenido en carnero. Conclusiones: Este trabajo permitió identificar y determinar la respuesta anticlases mayores de inmunoglobulinas en la muestra de estudio(AU)

Introduction: Immunoelectrophoresis is a precipitation technique that allows the characterization of complex biological samples. The Immunology Department of the Institute of Basic and Pre-Clinical Sciences Victoria de Girón has a hyperimmune antiserum obtained by immunization of sheep against human serum total proteins and it also has an anti-human IgA antiserum obtained from human colostrum. Objective: The aim of this study was to identify IgG, IgM and IgA in human serum and to determine response to anti-human IgM in human colostral IgA with antiserum obtained in sheep. Material and Methods: An observational descriptive cross-sectional study was conducted from November 2017 to June 2018. Immunoelectrophoresis of normal human serum was performed using hyperimmune antiserum. Results: These procedures allowed to identify IgG, IgM and IgA in addition to albumin and other protein fractions and to determine response to anti-human IgM in human colostral IgA with antiserum obtained in sheep. Conclusions: This work allowed us to identify and determine significant anti-class responses of immunoglobulins in the sample studied(AU)

Flucloxacillin-Induced Hepatotoxicity - Association with HLA-B*5701

SUMMARY Drug-induced liver injury (DILI) to flucloxacillin is rare and is classified as idiosyncratic, as it is dependent on individual susceptibility, unpredictable, and dose-independent. The authors present the case of a 74 - year - old man with a history of monoclonal gammopathy under investigation and alcoholic habits of 24 g/day, with asthenia, anorexia, nausea, abdominal discomfort, and fever with three days of evolution. He was treated with two courses of antibiotic therapy with flucloxacillin to erysipelas previously (3 months and 2 weeks before admission). Lab tests showed serum AST levels of 349 U/L, ALT 646 U/L, alkaline phosphatase 302 U/L, GGT 652 U/L, total bilirubin 3.3 mg/dL and direct bilirubin 2.72 mg/dL. Infectious, autoimmune, and metabolic causes were ruled out. Magnetic resonance cholangiopancreatography showed normal results. Liver biopsy showed mild multifocal (predominantly microvesicular) steatosis; marked changes in the centrilobular areas (sinusoidal dilatation, marked congestion, hemorrhage, and multifocal hepatocyte collapse); expansion of the portal areas with the formation of bridges; proliferated bile ducts and inflammatory infiltrate of variable density, predominantly mononuclear type. The HLA-B*5701 screening test was positive. Hepatic biochemical tests remain abnormal with a significative increase in total bilirubin, which reached levels of 24.1 mg/dL, with the development of jaundice, pruritus, and choluria. DILI was assumed, and the patient was treated with ursodeoxycholic acid. There was favorable evolution, without evidence of blood coagulation dysfunction or encephalopathy. The analytic normalization was, however, slow, with evolution to chronicity. The authors present this case to remind the possibility of moderate/severe drug-induced liver injury to flucloxacillin, an antibiotic commonly used in clinical practice and association with the HLA-B * 5701 allele reported in the literature.

RESUMO A hepatotoxicidade à flucloxacilina é rara e classifica-se como idiossincrática, uma vez que é dependente da suscetibilidade individual, não expectável e independente da dose. Apresentamos o caso de um homem, 74 anos, antecedentes de gamapatia monoclonal e hábitos alcoólicos de 24 g/dia, com quadro de astenia, anorexia, náuseas, desconforto abdominal e febrícula com três dias de evolução. Referência a dois ciclos de antibioterapia com flucloxacilina por erisipela (três meses e duas semanas antes da admissão). Analiticamente com AST 349 U/L, ALT 646 U/L, FA 302 U/L, GGT 652 U/L, bilirrubina total 3,3 mg/dL, bilirrubina direta 2,72 mg/dL. Excluídas etiologias infecciosa, autoimune, metabólica, bem como patologia das vias biliares por colangio-RM. Biópsia hepática mostrou esteatose multifocal ligeira (predominantemente microvesicular); alterações acentuadas nas áreas centrolobulares (dilatação sinusoidal, congestão acentuada, hemorragia e colapso multifocal de hepatócitos); expansão das áreas portais com constituição de pontes; ductos biliares proliferados e infiltrado inflamatório de densidade variável, predominantemente de tipo mononucleado. Tipagem de HLA-B*5701 positiva. Agravamento analítico atingindo bilirrubina total 24,1 mg/dL, com desenvolvimento de icterícia, prurido e colúria. Admitida a hepatotoxicidade, iniciou terapêutica com ácido ursodesoxicólico. Verificou-se evolução favorável, sem evidência de coagulopatia ou encefalopatia. A normalização analítica foi, no entanto, lenta, com evolução para cronicidade. Os autores apresentam este caso para alertar para a possibilidade de hepatotoxicidade moderada a grave à flucloxacilina, antibiótico de uso comum na prática clínica e associação com o alelo HLA-B*5701 relatada na literatura.

Necrobiotic Xanthogranuloma Coexists with Diffuse Normolipidemic Plane Xanthoma and Multiple Myeloma

Interferencia por iopamidol en el proteinograma por elecroforesis capilar y su relación con la función renal / Iopamidol interference in protein separation by capillary electrophoresis and its connection to renal function

Introducción: Las interferencias en el proteinograma electroforético por electroforesis capilar incluyen la aparición de picos con concentraciones y movilidades electroforéticas, que podrían simular la presencia de un componente monoclonal. Objetivos: Ante la aparición de un pico adicional con movilidad intera2-ß por electroforesis capilar (Minicap®-Sebia), el objetivo fue identificar el interferente y evaluar su relación con la funcionalidad renal. Material y métodos: Se estudiaron muestras de suero que presentaron dicha interferencia en un período de un año mediante proteinograma en soporte sólido, electroinmunofijación e inmunoelectroforesis. Se adicionó in vitro el probable interferente para confirmar su movilidad electroforética. Se evaluó el impacto de la corrección de la interferencia con la herramienta "eliminación de artefactos" (Phoresis®-Sebia) y la correlación de la concentración del pico a línea de base del interferente con la estimación de la tasa de filtrado glomerular (CKD- EPI). Resultados: La integración a la línea de base de los picos fue de 0,07-0,36 g/dL. No se observaron particularidades al realizar los estudios complementarios. Se evidenció, en todos los casos, la administración de iopamidol como medio de contraste, confirmándose su movilidad electroforética por su adición in vitro. Mediante la herramienta "eliminación de artefactos" se recuperaron los niveles basales de las fracciones. Se demostró la existencia de una correlación entre la concentración del pico a línea de base del interferente y la estimación de la tasa de filtración glomerular por CKD-EPI (r=-0.534, p<0.0001). Conclusiones: Se identificó al interferente como Iopamidol y se demostró su relación con la disminución de la tasa de filtración glomerular

Introduction: Interferences in the electrophoretic proteinogram by capillary electrophoresis include the appearance of peaks with concentrations and electrophoretic mobilities, which could simulate the presence of a monoclonal component. Objectives: In the light of an additional peak with interα2-ß mobility by capillary electrophoresis (MINICAP®-Sebia), the aim was to identify the interferent and evaluate its connection to renal functionality. Methods: Serum samples that presented this interference over a period of one year were studied by proteinogram on solid support, electroimmunofixation and immunoelectrophoresis. The probable interferent was added in vitro to confirm its electrophoretic mobility. The impact of the interference correction with the "artifact removal" tool (Phoresis®-Sebia) and the correlation of the baseline peak concentration of the interferent with the estimation of the glomerular filtration rate (CKD-EPI) were evaluated. Results: The integration to the baseline of the peaks was 0.07-0.36 g/dL. No particularities were observed when performing the complementary studies. In all cases, the administration of Iopamidol as a contrast medium was demonstrated, confirming its electrophoretic mobility due to its in vitro addition. Using the "artifact removal" tool, the basal levels of the fractions were recovered. The existence of a correlation between the concentration of the baseline peak of the interferent and the estimation of the glomerular filtration rate by CKD-EPI was shown (r=-0.534, p <0.0001). Conclusions: The interferent was identified as Iopamidol and its connection to the decrease in the glomerular filtration rate was demonstrated

Fibrillary glomerulonephritis combined with chronic inflammatory demyelinating polyneuropathy

A 58-yr-old man presented with leg edema and subacute weakness of his bilateral lower extremities. Urinary and serum immunoelectrophoresis revealed the presence of lambda-type Bence Jones proteins. He was ultimately diagnosed with monoclonal gammopathy of undetermined significance (MGUS). A renal biopsy specimen showed fibrillary glomerulonephritis (FGN), which was randomly arranged as 12-20 m nonbranching fibrils in the basement membranes. Immunofluorescence studies were negative for immunoglobulin (Ig)G, IgM, IgA, C3, and kappa light chains in the capillary walls and mesangial areas. A Congo red stain for amyloid was negative. Electromyography and nerve conduction velocity examinations results were compatible with the presence of demyelinating polyneuropathy. This case showed a rare combination of FGN, without Ig deposition, and MGUS combined with chronic inflammatory demyelinating polyneuropathy (CIDP).

Whole-cell antigenicity data support sequence-based kinetoplastid taxonomy

Early immunological data, obtained by immunodiffusion and immunoelectrophoresis, on the whole-cell antigenicity of kinetoplastid protozoa were retrieved and used to construct a dendrogram of antigenic distances. Remarkably, they supported the same taxonomic conclusions as analyses based on DNA and protein sequence data.

Establishment of a purificatory method for alpha-fetoprotein variant by affinity adsorption / 中华实验和临床病毒学杂志

<p><b>OBJECTIVE</b>To establish a purificatory method of alpha-fetoprotein variant (AFP-L3) based on microspincolumn with lens culinaris agglutinin (LCA).</p><p><b>METHODS</b>LCA was isolated by ammonium sulfate precipitation method from lens culinaris. AFP-L3 affinity adsorption microspincolumns which were made from LCA coupled with activated Sepharose 4B were prepared. By adding into the centrifuge column, serum was absorbed and eluted to purify AFP-L3. The results of purified AFP-L3 detection of 10 cases AFP positive sera by electro-chemiluminescence immunoassay were compared with traditional crossed affinity immunoelectrophoresis.</p><p><b>RESULTS</b>8 of 10 cases AFP-L3 concentration were greater than 5 ng/ml in purified sera. Six cases show positive reaction in affinity immune cross electrophoresis experiment.</p><p><b>CONCLUSION</b>Successfully established purification method of AFP-L3 by affinity absorption based on microspincolumn. The method was more conducive to clinical laboratory applications due to its high sensitive and easy operation.</p>

Multiple Myeloma with Biclonal Gammopathy Accompanied by Prostate Cancer / 대한진단검사의학회지

We report a rare case of multiple myeloma with biclonal gammopathy (IgG kappa and IgA lambda type) in a 58-year-old man with prostate cancer who presented with lower back pain. Through computed tomography (CT) imaging, an osteolytic lesion at the L3 vertebra and an enhancing lesion of the prostate gland with multiple lymphadenopathies were found. In the whole body positron emission tomography-computed tomography (PET-CT), an additional osteoblastic bone lesion was found in the left ischial bone. A prostate biopsy was performed, and adenocarcinoma was confirmed. Decompression surgery of the L3 vertebra was conducted, and the pathologic result indicated that the lesion was a plasma cell neoplasm. Immunofixation electrophoresis showed the presence of biclonal gammopathy (IgG kappa and IgA lambda). Bone marrow plasma cells (CD138 positive cells) comprised 7.2% of nucleated cells and showed kappa positivity. We started radiation therapy for the L3 vertebra lesion, with a total dose of 3,940 cGy, and androgen deprivation therapy as treatment for the prostate cancer.

Effect of Omega-3 Fatty Acids on Low Density Lipoprotein Subfraction, Adiponectin and Apolipoprotein B in Type 2 Diabetic Patients

BACKGROUND: Omega-3 fatty acids derived from fish oil have been reported to exert a beneficial effect on reducing cardiovascular disease. Reports about their mechanism have generated several interesting findings, including a change in small dense low density lipoprotein (sdLDL) cholesterol proportion, adiponectin, and apolipoprotein B (apoB), in addition to changes in the lipid profile. The principal objective of our study was to evaluate the effects of omega-3 fatty acids on plasma sdLDL, adiponectin, apoB100, and B48 in type 2 diabetic patients with hypertriglyceridemia. METHODS: We randomized 28 type 2 diabetic patients in a placebo-controlled, double-blind trial to receive either omega-3 fatty acids or placebo, both administered at a dose of 4 g daily for 12 weeks. LDL subfractions prior to and after treatment were separated via low-speed ultracentrifugation and analyzed via immunoelectrophoresis. Adiponectin, apoB100, and B48 levels were measured using an ELISA kit. RESULTS: sdLDL proportions were reduced in the omega-3 fatty acids group by 11% after 12 weeks of treatment (n = 17, P = 0.001), and were reduced by 4% in the control group (n = 11, P = 0.096). The patients receiving the omega-3 fatty acids evidenced a significant reduction in the levels of triglyceride (P = 0.001), apoB100, and B48 after 12 weeks (P = 0.038 and P = 0.009, respectively) relative to the baseline. Omega-3 fatty acids supplementation increased fasting blood glucose (P = 0.011), but the levels of HbA1c in each group did not change to a statistically significance degree. The adiponectin value was not reduced in the omega-3 fatty acids group (P = 0.133); by way of contrast, the placebo group evidenced a significant reduction in adiponectin value after 12 weeks (P = 0.002). CONCLUSION: Omega-3 fatty acid treatment proved effective in the reduction of atherogenic sdLDL and apoB in type 2 diabetic patients (Clinical trials reg. no. NCT 00758927, clinicaltrials.gov).

Systemic Plasmacytosis: A Case Report with a Review of the Literature

Systemic plasmacytosis is an uncommon disorder characterized by widely disseminated macular skin eruptions composed of polyclonal lymphoplasmacytic infiltrates associated with variable extracutaneous involvement. An aggressive clinical course has been observed in a small number of patients, but most cases have followed chronic and benign clinical course without spontaneous remission. Previously reported cases of this entity have been described almost exclusively in Japanese patients. We recently experienced a case of systemic plasmacytosis in a 48-year-old Korean female patient. Initial skin biopsy specimen revealed patchy perivascular and periadnexal infiltrates of mature plasma cells. Serum immunoelectrophoresis revealed polyclonal hypergammaglobulinemia, and polyclonal plasmacytosis was noted on the subsequent biopsy specimens of left supraclavicular and axillary lymph nodes. Multiple tiny pulmonary nodules appeared six years after the initial cutaneous presentation and were found to be of the same histologic appearance. We herein report a rare case of systemic plasmacytosis with a review of the literature.

A Case of Central Nervous System Myelomatosis with Complex Chromosome Aberrations / 대한진단검사의학회지

Involvement of the central nervous system is very uncommon in multiple myeloma, observed in approximately 1% of the multiple myeloma patients. We report a case of central nervous system myelomatosis with complex chromosome aberrations in a 62-yr-old female patient, who had previously been diagnosed as multiple myeloma. Fluorescent in situ hybridization revealed 13q deletion, p53 gene deletion and IGH/FGFR3 rearrangement and chromosomal study showed complex chromosome aberrations. After four cycles of chemotherapy, the patient was admitted to the hematology department with severe headache. Plasma cells were found in the cerebrospinal fluid (CSF), and CSF immunoelectrophoresis revealed abnormal precipitin arcs against anti-IgG and anti-lambda antisera. She was given systemic chemotherapy and eight courses of intrathecal chemotherapy, which cleared plasma cells in the CSF. Two months later, she was given autologous stem cell transplantation. Three months after stem cell transplantation, central nervous system myelomatosis progressed to plasma cell leukemia and two months later,the patient expired.

[Study of the immunological cross-reactions between goats and cows milk proteins]

Immunoelectrophoresis analysis showed immunological cross reactions between goat and cow milk caseins which belong to beta-casein, however, no such reaction were observed between goat and cow beta-lactoglobulin and alpha-lactalbumin. Systemic anaphylaxis test in guinea pigs showed strong immunological reactions between goat and cow milk proteins, injection of cow milk in animal's vein, which fed on cow milk caused 100% mortality. Same results were also obtained with injection of goat milk. Passive hemagglutination test against goat and cow milk was used to estimate antibody titer in guinea pigs serum, which fed cow's milk. The obtained results showed that the highest titer was found against casein followed by beta-lactoglobulin and alpha-lactalbumin for cow milk proteins, while for goat milk proteins the highest titer was found against casein followed by alpha-lactalbumm and beta-lactoglobulin. The titer of antibodies against goat alpha[s]-casein and Kappa-casein was lower than that for cow milk, the behavior of goat and cow beta-casein was similar for both proteins

Clinical research of AFP variant with microcentrifugalcolumn method and crossed affinity immunoelectrophoresis autoradiography method / 中华实验和临床病毒学杂志

<p><b>OBJECTIVE</b>To compare the clinical value of microcentrifugalcolumn method with crossed affinity immunoelectrophoresis autoradiography method for the measurement of alpha-fetoprotein variant (AFP-L3) in differentiation of benign and malignant liver.</p><p><b>METHODS</b>Serum AFP-L3 variants in 102 primary hepatocellular carcinoma patients and 41 chronic and cirrhosis patients were separated by microcentrifugalcolumn method and crossed affinity immunoelectrophoresis autoradiography method and the clinical value of both method were compared.</p><p><b>RESULTS</b>In 102 primary hepatocellular carcinoma patients, the sensitivity of AFP-L3 was 79.4% and 91.2% respectively, in 41 chronic and cirrhosis patients. The specificity of AFP-L3 was 70.7% and 29.3% respectively. The diagnostic accuracy was 76.9% and 73.4% respectively. The area in the ROC curve was 0.791 and 0.758 respectively. In the 4 primary hepatocellular carcinoma patients with lower AFP-L3, the AFP-L3 was positive by useing microcentrifugalcolumn method, but none of AFP-L3 were found by useing crossed affinity immunoelectrophoresis autoradiography method.</p><p><b>CONCLUSION</b>The micro centrifugalcolumn method is more simple and rapid, it may be more useful in differentiation of benign and malignant liver than traditional crossed affinity immunoelectrophoresis autoradiography method.</p>

Serum Free Light Chains for Diagnosis and Follow-up of Multiple Myeloma / 대한진단검사의학회지

BACKGROUND: Free light chain (FLC) is widely used to evaluate B-cell proliferative diseases. Herein, we estimated the clinical usefulness of serum FLC in multiple myeloma (MM). METHODS: Fifty-one patients were enrolled. We performed FLC analysis, protein electrophoresis (PEP), and immunofixation electrophoresis (IFE). FLC was measured using Toshiba 200 FR Neo with FREELITE(TM), and kappa/lambda (kappa/lambda) ratio was calculated. We compared these parameters in 41 patients with increased FLC before and after bortezomib treatment. Complete response (CR) was defined as the disappearance of monoclonal (M) protein in serum and/or urine as measured by IFE. Partial response (PR) was defined as > or =50% reduction of serum M protein. Early objective response (EOR) included both CR and PR. Minimal response (MR) was defined as 25-49% reduction of M protein and stable disease (SD) as <25% reduction. RESULTS: Forty-one (80.4%) of the 51 patients studied revealed increment of FLC and the five patients with no increment revealed an abnormal kappa/lambda ratio. Especially, all of the light chain myeloma and non-secretory myeloma showed increased FLC concentrations. Among the patients with EOR, 72.4% (21/29) showed a normal or subnormal FLC concentration after the first cycle of treatment. Otherwise, PEP and IFE normalized in 24.1% (7/29) and 24.1% (7/29), respectively. The ratio of decreased FLC after the first cycle of treatment was significantly different between EOR and other response groups (MR, SD) (90.6% vs 51.8%, P=0.011). CONCLUSIONS: FLC was considered as a good diagnostic method in complement with PEP and IFE in MM, especially in light chain myeloma or non-secretory myeloma. Moreover, FLC is a useful monitoring tool because it reflects therapy results more rapidly owing to a short serum half-life.

Immunoglobulin heavy and light chain isotypes in multiple myeloma patients.

The frequency of expression of immunoglobulin (Ig) light and heavy chain isotypes was analyzed in myeloma proteins (M-proteins) from sera of 40 Indian patients with clinically established multiple myeloma. Patients samples were screened by a combination of electrophoresis, immunoelectrophoresis (IEP) and ELISA techniques in this study. We found that majority of the myeloma proteins (58%) were of the IgG isotype followed by IgA (24%) and biclonal gammopathy associated with IgG and IgA (5%). Both kappa and lambda light chains were associated with the heavy chain isotypes. We recommend the triangular combination for detection of M-proteins and biclonal gammopathy of cancerous plasma cells as biomarkers for diagnosis of myeloma.

Inhibition of enterotoxigenic Escherichia coli (ETEC) adhesion to Caco-2 cells by human milk and its immunoglobulin and non-immunoglobulin fractions

Enterotoxigenic Escherichia coli (ETEC) is the most common cause of diarrhea in children in developing countries and among travelers to ETEC endemic areas. ETEC diarrhea is caused by colonization of the small intestine mediated by colonization factor (CF) antigens, and subsequent elaboration of enterotoxins. Breast feeding has been related to protection against enteric infections. The protective effect of human milk can be ascribed to its immunoglobulin content, specially secretory immunoglobulin A (sIgA), and to non-immunoglobulin components such as free oligosaccharides, glycoproteins and glycolipids. In this study we investigated the effect of whole human milk and its fractions immunoglobulin and non-immunoglobulin on the adherence of ETEC strains possessing different CFs to Caco-2 cells, as well as the ability of sIgA and free secretory component (fSC) to bind to bacterial superficial proteins. Pooled human milk from three donors were fractionated by gel filtration and analyzed by SDS-PAGE. Our results revealed that whole human milk and its proteins fractions, containing sIgA and fSC, inhibited adhesion ETEC strains harboring different colonization factors antigens. We also verified that sIgA and fSC, using immunoblotting and immunogold labeling assays, bound to some fimbrial proteins and other material present in bacterial surface. Our findings suggest that whole human milk and its fractions may contribute to protection against ETEC infections by blocking bacterial adhesion mediated by different colonization antigens.

Escherichia coli enterotoxigênica (ETEC) é uma importante causa de diarréia infantil em países em desenvolvimento e entre os viajantes em áreas endêmicas. A diarréia por ETEC é causada pela colonização do intestino delgado, mediada pelos fatores antigênicos de colonização (CFs), e a subseqüente elaboração de enterotoxinas. A amamentação tem sido relacionada com a proteção contra infecções entéricas. O efeito protetor do leite humano pode ser atribuído ao seu conteúdo imunoglobulínico, especialmente a imunoglobulina A secretória (sIgA), e aos componentes não imunoglobulínicos tais como oligossacarídeos livres, glicoproteínas e glicolipídeos. Neste estudo nós investigamos o efeito do leite humano total e suas frações imunoglobulínica e não imunoglobulínica sobre a adesão de cepas ETEC possuidoras de diferentes fatores de colonização as células Caco-2, bem como a habilidade da sIgA e do componente secretor livre (fSC) de se ligarem as proteínas da superfície bacteriana. Uma mistura de leite de três doadoras foi fracionada por filtração em gel e analisada por SDS-PAGE. Nossos resultados revelaram que o leite humano total e suas frações protéicas, contendo sIgA e fSC, inibiu a adesão de cepas ETEC albergando diferentes fatores antigênicos de colonização. Nós também verificamos que a sIgA e o fSC, usando ensaios de imunoeletrotransferência e imunomarcação, ligaram-se a algumas proteínas fimbriais e a outro material presente na superfície bacteriana. Nossos dados sugerem que o leite humano total e suas frações podem contribuir para proteção contra as infecções por ETEC bloqueando a adesão bacteriana mediada pelos diferentes antígenos de colonização.

Testes laboratoriais para avaliação do componente monoclonal / Laboratory tests for evaluating the M-component

As gamopatias monoclonais resultam de hiperprodução de um único clone anormal de células plasmocitárias ou linfócitos B. O objetivo da avaliação laboratorial nas gamopatias é demonstrar a presença, a quantidade e o tipo de proteína anormal presente no soro e/ou na urina através do estudo do perfil protéico, quantificação das imunoglobulinas e cadeias leves e avaliação da proteinúria. Este artigo descreve as principais técnicas laboratoriais disponíveis, bem como suas indicações e limitações.

Monoclonal gammopathies result from an overproduction of a single abnormal clone of a plasma cell or B lymphocyte. The purpose of the laboratory protocols in these situations is to demonstrate the presence, the characterization and the concentration of an abnormal protein detected in serum and/or urine samples. The laboratory investigation is based on the electrophoretic protein profile, quantification of immunoglobulins, free light chains and proteinuria. This paper describes the major available laboratory methods as well their indications and limitations.

A Case of Solitary Bone Plasmacytoma Presenting with Erythematous Swelling of the Upper Eyelid

PURPOSE: To demonstrate a rare case of orbital solitary bone plasmacytoma that presented with erythematous swelling of the upper eyelid. METHODS: A 64-year-old woman presented with a erythematous swelling of the right upper eyelid that persisted for 8 months prior to examination. There were 3 mm of proptosis in the right eye on ophthalmologic examination and an orbital tumor was detected on computed tomography (CT) scan, infiltrating the intracranial space and the sphenoid bone. Therefore, we performed incisional biopsy through the right upper eyelid. RESULTS: Histopathologic examination revealed the tumor to be a plasmacytoma (lambda light chain-type) and all the systemic work-ups (serologic and urinary protein immunoelectrophoresis, bone marrow biopsy, radiologic examination) to rule out multiple myeloma were negative. Because of these factors, a diagnosis of solitary bone plasmacytoma was made. Radiation therapy (5,000 Gy/25 fractions) was performed over 5 weeks and the size of the tumor was found to have markedly decreased on a follow-up CT scan. CONCLUSIONS: Solitary bone plasmacytoma is rare and has variable prognoses. This condition should be considered when the presenting symptom is eyelid swelling and proptosis.

Rabbit IgG antibodies against Phospholipase A2 from Crotalus durissus terrificus neutralize the lethal activity of the venom

Crotalus durissus terrificus (C.d.t.) (South American rattlesnake) venom possesses myotoxic and neurotoxic activities, both of which are also expressed by crotoxin, the principal toxin of this venom. Crotoxin contains a basic phospholipase A2 (PLA2) and a non toxic acidic protein, crotapotin. We have produced and investigated the ability of IgG antibodies raised in rabbits against PLA2 to neutralize the lethality of the whole venom. PLA2 was isolated by gel filtration chromatography (Sephadex G-75). Specific antibodies were obtained by subcutaneous and intramuscular inoculation of PLA2 (700 µg) with Freund adjuvant. Groups of six mice (20 + 2 g) were inoculated with 0.5 ml i.p. of C. d. t. venom (4 µg) or a mixture of venom that had been preincubated with the desired volume of IgG antibodies. Mortality, recorded 24 and 48 h after inoculation, showed that IgG anti-PLA2 were more effective than anticrotalic serum in neutralizing the lethal activity. These results demonstrate that it could be possible to obtain an anti-venom made by specific antibodies with a high level of protection against the lethal component of C.d.t. venom, and/or the inclusion of these antibodies as a supplement in heterologous anti-venoms

El veneno de Crotalus durissus terrificus (C.d.t.) (Cascabel de Sud América) posee actividad miotóxica y neurotóxica, actividades que también exhibe el complejo crotoxina, principal componente tóxico de este veneno. El complejo crotoxina está constituido por una fosfolipasa A2 básica (PLA2) y una proteína acídica no tóxica, el crotapotín. En este trabajo se estudió la capacidad neutralizante de anticuerpos IgG anti-PLA2 sobre la letalidad inducida por el veneno entero. El antígeno PLA2, fue aislado por cromatografía de filtración en gel (Sephadex G-75). Se inocularon conejos machos por vía subcutánea e intramuscular, con 700 µg de PLA2 y adyuvante para la obtención de anticuerpos específicos. La capacidad neutralizante del antisuero se analizó en ratones por inoculación con diluciones de veneno entero preincubado con un volumen adecuado de anticuerpos IgG anti-PLA2. Se inocularon ratones controles con 0.5 ml i.p. de veneno (4 µg.ml-1). El número de muertes fue contabilizado a las 24 y 48 h posteriores a la inoculación, demostrándose que la capacidad neutralizante de los anticuerpos IgG anti-PLA2 fue superior a la obtenida con el antiveneno crotálico. Los resultados obtenidos demuestran la potencial aplicación de antivenenos constituidos por anticuerpos específicos contra PLA2, y/o la inclusión de estos anticuerpos como suplementos en antivenenos polivalentes