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1.
Journal of Experimental Hematology ; (6): 1209-1215, 2021.
Article in Chinese | WPRIM | ID: wpr-888540

ABSTRACT

OBJECTIVE@#To investigate the comparability of the Freelite, Binding Site, Beckman and N Latex FLC, Siemens in the detection of serum free light chain (sFLC) .@*METHODS@#Fifty newly diagnosed multiple myeloma (MM) patients in Tianjin Institute of Blood Research from November 2019 to February 2020 were enrolled. The two systems (Freelite, Binding Site, Beckman and N Latex FLC, Siemens) were used to detect the sFLC of the samples. Outlier detection was performed by ESD method, methodological comparison and deviation assessment were performed by Passing-Bablok regression and Bland-Altman regression.@*RESULTS@#Both the systems could quantitatively analyze free kappa light chain serum samples and free lambda light chain samples. Freelite, Binding Site, Beckman and N Latex FLC, Siemens free light chain test showed FLC-κ:36.5 (6.5, 194), 40.5 (6.94, 288), FLC-λ: 30.1 (4.3, 170.5), 35.1 (2.28, 526), rFLC (FLC-κ/ FLC-λ) : 0.82 (0.05, 43.25), 1.03 (0.03, 32.04), dFLC (|FLC-κ- FLC-λ|) : -5.8 (-161.97, 183.7), 1.1 (-505.1, 279.01), which existed no outliers. There were systematic differences, and the deviation level was not within the clinically acceptable range.@*CONCLUSION@#Both the systems can meet the needs of clinical diagnosis and treatment, but there is a significant deviation between the two systems, the results are not comparable, and should be analyzed separately. In particular, the same system should be selected for monitoring the prognosis of MM.


Subject(s)
Humans , Immunoglobulin Light Chains , Immunoglobulin kappa-Chains , Immunoglobulin lambda-Chains , Latex , Multiple Myeloma/diagnosis
2.
Article in Chinese | WPRIM | ID: wpr-827190

ABSTRACT

OBJECTIVE@#To explore the clinical application value of serum light chain (sLC) in the diagnosis and therapeutic efficacy evaluation for multiple myeloma.@*METHODS@#46 patients with newly diagnosed multiple myeloma were selected as MM group and 50 healthy persons as control group. Rate scattering immunoturbidimetry was used to detect serum light chain and immunoglobulin (Ig) in two groups, serum protein electrophoresis was used to detect M protein by agarose gel. Then, the sensitivity and specificity of the two methods in MM diagnosis were analyzed and compared, and the significance of sLC detection in MM diagnosis were discussed. In addition, 15 MM patients after received conventional therapy were tracked, sLC levels in five different therapentic times were recorded, and the effect of sLC in efficacy evaluation of MM was analyzed.@*RESULTS@#There were 11 cases of IgA type, 15 cases of IgG type, 8 cases of light chain κ type, 8 cases of light chain λ type, 2 cases of IgD type, and 2 cases of non-secretion type. The sLC-κ, sLC-λ and their ratio (including light chain type and double clone type), IgA and IgG (except IgD type), as well as albumin, beta-globulin and gamma-globulin levels showed statistically significant differences (P<0.05) compared with the control group. The sensitivity of serum protein electrophoresis, Ig quantification, sLC and its ratio in the diagnosis of multiple myeloma were 57%, 76% and 65%, and their specificity were 83%, 61% and 90%, respectively. After the second or third chemotherapy, the sLC-κ/λ ratio gradually approached the normal range as the disease reliefes, and the sLC-κ/λ ratio continued to be on or off the line at outliers or further away from the reference value as the disease progresses in MM patients with κ type or λ type.@*CONCLUSION@#sLC detection shows positive significance in early diagnosis of multiple myeloma, SLC monitoring can be used for the efficacy evaluation for treatment of MM patients.


Subject(s)
Humans , Immunoglobulin kappa-Chains , Immunoglobulin lambda-Chains , Multiple Myeloma , Diagnosis
3.
Journal of Experimental Hematology ; (6): 1173-1178, 2019.
Article in Chinese | WPRIM | ID: wpr-775746

ABSTRACT

OBJECTIVE@#To analyze the clinical significance of serum free light chain (sFLC) and sFLC-κ/λ ratio in patients with newly diagnosed multiple myeloma (MM), so as to provide the theoretical basis for the diagnosis, treatment and prognosis of MM patients.@*METHODS@#A total of 43 patients newly diagnosed as MM in our hospital and 40 cases of other diseases that could cause sFLC increase were selcted. The clinical features of newly diagnosed MM patients were analyzed. On the basis of the type of sFLC, MM patients were divided into sFLC-κ and sFLC-λ group. The correlation of sFLC with clinical index in two groups was analyzed. The difference of sFLC-κ/λ ratio between MM and non-MM patients was analyzed. According to whether the involved/uninvolved sFLC ratio was greater than 100, the MM patients were divided into the low ratio and the high ratio group. The clinical index of these 2 groups were analyzed.@*RESULTS@#Among MM patients, IgG type was the most. In sFLC-κ and -λ groups, the hemoglobin level negatively correlated with sFLC (r=-0.307, r=-0.494, P<0.05), and the serum β2 microglobulin level positively correlated with sFLC (r=0.453, r=0.689, P<0.05), and myeloma cells ratio also positively correlated with sFLC (r=0.307, r=0.309, P<0.05). The concentration of albumin, lactate dehydrogenase and serum calcium did not correlate with sFLC (P>0.05). The serum creatinine level also did not correlate with sFLC in the sFLC-κ group (P>0.05), but in the sFLC-λ group they were obviously positive (r=0.697, P<0.01). The ratio of sFLC-κ/λ <0.26 or >1.65 in MM patients was higher than that of non-MM patients, and the ratio of involved/uninvolved sFLC≥100 in MM patients was also higher than that of non-MM patients with statistically significant differences (P<0.01). The levels of serum albumin, creatinine and β2 microglobulin, as well as myeloma cell ratio and ISS international staging of the patients between 2 groups were significantly different (P<0.05), while the differences in sex, age and levels of hemoglobin, lactate dehydrogenase and blood calcium were not significant between the 2 groups (P>0.05).@*CONCLUSION@#The light chain type λ in MM patients correlates with renal damage. The abnormalities of sFLC-κ/λ ratio occur mostly in MM patients. The MM patients with involved/uninvolosed sFLC ratio ≥100 show the severe symptoms and poor prognosis.


Subject(s)
Humans , Immunoglobulin Light Chains , Immunoglobulin kappa-Chains , Immunoglobulin lambda-Chains , L-Lactate Dehydrogenase , Multiple Myeloma , Prognosis
4.
Egyptian Journal of Hospital Medicine [The]. 2018; 71 (2): 2556-2563
in English | IMEMR | ID: emr-192498

ABSTRACT

Background: kappa and lambda light chains detection in bone marrow trephine sections help in the determination of B-cell clonality through evaluation of light chain restriction


Aim of the Work: was to compare the efficacy of single color detection-based immunohistochemistry [IHC] and chromogenic in situ hybridization [CISH] in evaluating kappa/lambda expression in tissues harboring B-lymphoid lesions


Patients and Methods: Forty patients were enrolled in this study. They were divided into three groups chronic lymphocytic leukemia [CLL/SLL] group I [n=13], non-Hodgkin lymphoma [NHL] group II [n=24] and hairy cell leukemia [HCL] group III [n=3]. The 24 NHL cases comprised of [11 diffuse large B-cell lymphoma, 6 mantle cell lymphomas, 3 marginal zone lymphoma, 2 lymphoplasmacytic lymphoma, 1 follicular lymphomas and 1 Burkitt's lymphoma]. Kappa and lambda light chains were detected in their bone marrow trephine sections using single colored immunohistochemistry, chromogenic in situ hybridization and the results were compared to the flowcytometry as reference method


Results: Light chain restriction [LCR] was detected by FCM in 100% of the cases followed by CISH [52.1%; 12/23] of the cases and finally IHC [43%; 18/40]


Conclusion: Both conventional CISH and IHC are effective in determining monoclonality in cases of mature B- cell neoplasm that has plasmacytic differentiation and with high amount of cytoplasmic Ig light chains such as MZL and LP. However, they are not effective in determining monoclonality in cases with low amount of Ig light chain such as cases of pregerminal and germinal center lymphoma. Yet, CISH is more informative than IHC due to the lack of background staining which allowed for greater discrimination between absence and presence of monoclonality


Subject(s)
Humans , Male , Female , Adult , Middle Aged , Aged , Immunoglobulin kappa-Chains , Immunoglobulin lambda-Chains , Lymphoma, B-Cell/immunology , Immunohistochemistry , Chromogenic Compounds , In Situ Hybridization/methods , Gene Rearrangement, B-Lymphocyte, Light Chain
5.
Chinese Journal of Pathology ; (12): 100-105, 2015.
Article in Chinese | WPRIM | ID: wpr-298143

ABSTRACT

<p><b>OBJECTIVE</b>To study the clinicopathologic features, immunophenotype and gene rearrangement of primary cutaneous diffuse large B-cell lymphoma, leg type (PCLBCL).</p><p><b>METHODS</b>Seven cases of PCLBCL were enrolled into the study. Clinicopathologic analysis, immunohistochemical staining and gene rearrangement for IgH and Igκ were undertaken in the study.</p><p><b>RESULTS</b>All the seven cases were male, and the median age was 72 years. Patients usually presented with multiple purple tumors, nodules, papules and infiltrative plaques. Two patients had a history of leg injury before onset, and one had mosquito bites. Histologically, the tumor involved the dermis and subcutis with dense and diffuse infiltrative pattern composing of centroblasts and/or immunoblasts. Immunohistochemical staining showed that seven cases (7/7) expressed CD20, six (6/6) expressed bcl-2, four (4/4) expressed MUM-1, four (4/5) expressed CD79a, four (4/5) expressed PAX-5 and four (4/6) expressed bcl-6, respectively. All cases did not express CD3ε, CD45RO, CD10 and CD30. IgH gene rearranged bands were detected in three (3/6) cases and Igκ was detected in one (1/5) case. Six of the seven cases died and the remaining patient, who was 44-year-old, was alive after 22 months of follow-up.</p><p><b>CONCLUSIONS</b>PCLBCL is rare, predominantly affects elderly male patients. PCLBCL has poor prognosis and high mortality, but younger patients seem to have better prognosis. Some cases had a history of trauma or mosquito bites. The relationship between the history and the onset of PCLBCL needs further evaluation.</p>


Subject(s)
Aged , Aged, 80 and over , Animals , Antigens, CD , Culicidae , Gene Rearrangement , Humans , Immunoglobulin Heavy Chains , Genetics , Immunoglobulin kappa-Chains , Genetics , Immunophenotyping , Insect Bites and Stings , Leg , Leg Injuries , Lymphoma, Large B-Cell, Diffuse , Genetics , Metabolism , Pathology , Male , Middle Aged , Prognosis , Proto-Oncogene Proteins c-bcl-6 , Metabolism , Skin Neoplasms , Genetics , Pathology
6.
Journal of Experimental Hematology ; (6): 1357-1361, 2015.
Article in Chinese | WPRIM | ID: wpr-274036

ABSTRACT

<p><b>OBJECTIVE</b>To investigate the clinical significance of serum free light chain (sFLC) detection in light chain multiple myeloma (LCMM).</p><p><b>METHODS</b>A total of 37 newly diagnosed LCMM patients were enrolled in this study, including 17 patients with k light chain type and 20 patients with λ light chain type, the sFLC and 24 hours urine light chain (ULC) were measured before and after chemotherapy. The correlation of sFLC level with ULC and renal impairment was analyzed.</p><p><b>RESULTS</b>All the patients displayed an abnormally increased level of sFLC at diagnosis wtih median value of 105.44 mg/L and 146.39 mg/L for k and λ light chain types, respectively. The sFLC did not correlate with ULC before and after chemotherapy. Among the 12 patients with very good partial remission and normal ULC level, the sFLC still was abnormally increased in 8 patients. Renal impairment was associated with the urine λ-type light chain, and the area under the ROC curve of urine λ light chain at diagnosis is 0.792 (P = 0.031).</p><p><b>CONCLUSION</b>All patients with LCMM show an abnormally increased level of sFLC at diagnosis. sFLC can be used to monitor the response to chemotherapy because it is more sensitive for analysis of therapeutic effect than urine λ light chain.</p>


Subject(s)
Humans , Immunoglobulin Light Chains , Blood , Immunoglobulin kappa-Chains , Blood , Immunoglobulin lambda-Chains , Blood , Multiple Myeloma , Blood , Drug Therapy , Renal Insufficiency
7.
Article in Korean | WPRIM | ID: wpr-153827

ABSTRACT

A 53-year-old woman was admitted with epigastric discomfort and weakness. Laboratory examination at admission showed mild anemia and proteinuria. Esophagogastroduodenoscopy revealed marked mucosal atrophy, diffuse nodularity and granular appearance with mucosal friability. Biopsy was performed on the antrum and body of the stomach. On the next day, the patient began to complain of severe dyspnea, and hypoxia was present on pulse oximetry. Therefore, emergency echocardiography was conducted and it showed restrictive cardiomyopathy along with thrombus in the left atrium. With time, heart failure was aggravated despite intensive management. The result of gastric biopsy revealed amyloid deposits which stained positively with Congo red. On immunohistochemistry study, kappa and lambda chain were present. In addition, kappa chain was significantly elevated in urine and serum on electrophoresis. Although the patient was finally diagnosed as having primary gastric amyloidosis with restrictive cardiomyopathy, her general condition rapidly deteriorated and died at 12th hospital day. When obscure gastric lesion is encountered, performing gastric biopsy is strongly recommended since it be primary gastric amyloidosis. Herein, we present an unusual case of primary gastric amyloidosis.


Subject(s)
Amyloidosis/complications , Endoscopy, Digestive System , Female , Heart Atria/diagnostic imaging , Heart Failure/complications , Humans , Immunoglobulin kappa-Chains/blood , Immunoglobulin lambda-Chains/blood , Immunohistochemistry , Magnetic Resonance Imaging , Middle Aged , Stomach Diseases/complications , Thrombosis/diagnosis , Tomography, X-Ray Computed , Ultrasonography
8.
Braz. j. infect. dis ; 18(6): 638-642, Nov-Dec/2014. tab, graf
Article in English | LILACS | ID: lil-730423

ABSTRACT

Hepatitis C virus (HCV) infects B-lymphocytes, provokes cellular dysfunction and causes lymphoproliferative diseases such as cryoglobulinemia and non-Hodgkin's B-cell lymphoma. In the present study, we investigated the serum levels of kappa and lambda free light chains (FLC) of immunoglobulins and the kappa/lambda FLC ratio in Brazilian patients with chronic HCV infection and cryoglobulinemia. We also analyzed the immunochemical composition of the cryoglobulins in these patients. Twenty-eight cryoglobulinemic HCV patients composed the target group, while 37 HCV patients without cryoglobulinemia were included as controls. The median levels of kappa and lambda FLC were higher in patients with cryoglobulinemia compared to controls (p = 0.001 and p = 0.003, respectively), but the kappa/lambda FLC ratio was similar in patients with and without cryoglobulinemia (p > 0.05). The median FLC ratio was higher in HCV patients presenting with advanced fibrosis of the liver compared to HCV patients without fibrosis (p = 0.004). Kappa and lambda FLC levels were strongly correlated with the IgA, IgG and IgM levels in the patients with cryoglobulinemia. In patients without cryoglobulinemia, the kappa FLC level was only correlated with the IgG level, whereas the lambda FLC were weakly correlated with the IgA, IgG and IgM levels. An immunochemical pattern of mixed cryoglobulins (MC), predominantly IgM, IgG, IgA and kappa light chain, was verified in these immune complexes. We concluded that HCV-infected patients presenting cryoglobulinemia have vigorous polyclonal B-lymphocyte activation due to chronic HCV infection and persistent immune stimulation.


Subject(s)
Female , Humans , Male , Middle Aged , Cryoglobulinemia/etiology , Cryoglobulins/analysis , Hepatitis C, Chronic/complications , Immunoglobulin kappa-Chains/blood , Immunoglobulin lambda-Chains/blood , Case-Control Studies , Hepatitis C, Chronic/blood , Immunohistochemistry , Immunoglobulin G/blood , Immunoglobulin M/blood
9.
Article in English | WPRIM | ID: wpr-84046

ABSTRACT

Fanconi syndrome (FS) is a rare condition that is characterized by defects in the proximal tubular function. A 48-year-old woman was admitted for evaluation of proteinuria. The patient showed normal anion gap acidosis, normoglycemic glycosuria, hypophosphatemia, and hypouricemia. Thus, her condition was compatible with FS. The M peak was found behind the beta globulin region in urine protein electrophoresis. Upon bone marrow examination, we found that 24% of cells were CD138+ plasma cells with kappa restriction. From a kidney biopsy, we found crystalline inclusions within proximal tubular epithelial cells. Thereafter, she was diagnosed with FS accompanied by multiple myeloma. The patient received chemotherapy and autologous stem cell transplantation, and obtained very good partial hematologic response. However, proximal tubular dysfunction was persistent until 1 year after autologous stem cell transplantation. In short, we report a case of FS accompanied by multiple myeloma, demonstrating crystalline inclusion in proximal tubular cells on kidney biopsy.


Subject(s)
Acid-Base Equilibrium , Acidosis , Beta-Globulins , Biopsy , Bone Marrow Examination , Crystallins , Drug Therapy , Electrophoresis , Epithelial Cells , Fanconi Syndrome , Female , Glycosuria , Humans , Hypophosphatemia , Immunoglobulin kappa-Chains , Kidney , Middle Aged , Multiple Myeloma , Plasma Cells , Proteinuria , Stem Cell Transplantation
10.
Article in English | WPRIM | ID: wpr-200212

ABSTRACT

A mediastinal mass was incidentally found on chest radiography in a 46-yr-old woman who had had myasthenia gravis (MG) for 2 months. Computed tomography revealed a 4-cm in size, well-defined, and lobulating mass with nodular calcification that was located in the thymus. Microscopically, the mass consisted of diffuse amorphous eosinophilic materials. These deposits exhibited apple-green birefringence under polarized light microscopy after Congo red staining. Immunohistochemical analysis revealed that they were positive for both kappa and lambda light chains and negative for amyloid A. A diagnosis of localized primary thymic amyloidosis was finally made. After thymectomy, the symptoms of MG were controlled with reduced corticosteroid requirements. Localized thymic amyloidosis associated with MG has not been reported to date.


Subject(s)
Amyloidosis/complications , Calcinosis/diagnostic imaging , Female , Humans , Immunoglobulin kappa-Chains/immunology , Immunoglobulin lambda-Chains/immunology , Mediastinum/diagnostic imaging , Middle Aged , Myasthenia Gravis/complications , Radiography, Thoracic , Thymectomy , Thymus Gland/diagnostic imaging , Tomography, X-Ray Computed
11.
Rev. cuba. hematol. inmunol. hemoter ; 29(2): 183-188, abr.-jun. 2013.
Article in Spanish | LILACS | ID: lil-672147

ABSTRACT

Introducción: el mieloma múltiple (MM) es una enfermedad caracterizada por una proliferación monoclonal de inmunoglobulinas que representa aproximadamente el 15 por ciento de las hemopatías malignas. Métodos: se realizó un estudio de la distribución de las clases, sub clases y tipos de cadenas ligeras de inmunoglobulinas en 285 enfermos con el diagnóstico de MM. Se emplearon tres métodos: electroforesis de proteínas en suero para la detección de la inmunoglobulina monoclonal o paraproteína, electroforesis de inmunofijación y doble inmunodifusión para identificar las clases, sub clases y tipo de cadenas ligeras. Resultados: se encontraron 206 enfermos (72.28 por ciento) con MM IgG; 73 (25.62 por ciento) con MM IgA y 6 (2.1 por ciento) con MM IgM. La distribución de sub clases de IgG fue: 130 casos (63.11 por ciento) IgG1, 43 (20.87 por ciento) IgG2, 21 (10.19 porciento) IgG3 y 12 (5.83 por ciento) IgG4; y la de sub clases de IgA fue de 59 enfermos (80.82 por ciento) IgA1 y 14 (19.18 por ciento) IgA2. Del total de enfermos 187 (65.61 por ciento) mostraron cadenas ligeras tipo kappa y 98 (34.38 por ciento) tipo lambda. Conclusiones: los datos obtenidos en nuestro estudio permitieron identificar la frecuencia de distribución de las clases, subclases y cadenas ligeras en una muestra de enfermos con MM


Introduction: multiple mieloma (MM) is a disease characterized by a monoclonal proliferation of immunoglobulins representing approximately 15 percent of malignant hemopathies. Methods: the distribution of classes, subclasses and light chains of monoclonal immunoglobulins was studied in 285 patients with MM. Three methods were used: serum protein electrophoresis for the detection of monoclonal immunoglobulins or paraproteins, immunofixation electrophoresis and double immunodiffusion to identify classes, subclasses and light chain types. Results: 206 patients (72.28 percent) with IgG MM, 73 (25,62 percent) with IgA MM, and 6 (2,1 percent) with IgM MM were found. The distribution of IgG subclasses was: 130 cases (63,11 percent) IgG1; 43 (20,87 percent) IgG2; 21 (10,19 percent) IgG3: and 12 (5,83 percent) IgG4. Distribution of IgA subclasses was: 59 patients (80,82 percent) IgA1 and 14 (19,18 percent) IgA2; 187 patients (65,62 percent) showed kappa light chains and 98 (34,38 percent) were lambda. Conclusions: the data obtained in our study allowed us to identify the frequency of distribution of classes, subclasses and light chains in a sample of patients with MM


Subject(s)
Hemoglobin A/analysis , Multiple Myeloma/complications , Paraproteins/analysis , Myeloma Proteins/analysis , Immunoglobulin kappa-Chains/analysis , Blood Protein Electrophoresis/methods , Electrophoresis/methods , Immunoglobulin Light Chains
12.
Article in Chinese | WPRIM | ID: wpr-306487

ABSTRACT

<p><b>OBJECTIVE</b>To construct full-length human bladder cancer-specific antibody libraries for efficient display of full-length antibodies on the surface of mammalian cells.</p><p><b>METHODS</b>The total RNA was isolated from peripheral blood mononuclear cells from patients with bladder cancer. The repertoires of IgG1 heavy chain variable region (VH) and Kappa light chain were amplified by RT-PCR using specific primers. The antibody genes were inserted into the vector pDGB-HC-TM to construct the bladder-cancer-specific antibody libraries of heavy chains and light chains. Ten clones from each library were randomly picked for gene sequencing and transient transfection into FCHO cells to analyze antibody display on mammalian cell surface by flow cytometry after staining with corresponding fluorescent labeled antibodies.</p><p><b>RESULTS</b>The libraries of bladder-cancer-specific antibody heavy chain (IgG1) and light chain (LCk) were successfully constructed. Seven out of the 10 clones randomly selected from the heavy chain library and 9 out of the 10 clones from the light chain library showed correct open reading frame, coding for 7 unique VH and 9 unique LCk. The combinatory library size reached 3.32×10(11).</p><p><b>CONCLUSION</b>We have successfully constructed a full-length human bladder-cancer-specific antibody library with a combinatory diversity of 3.32×10(11) based on mammalian display technology, which can be used for screening monoclonal antibodies against bladder-cancer-associated antigens.</p>


Subject(s)
Amino Acid Sequence , Animals , Antibodies , Genetics , Cell Surface Display Techniques , Gene Library , Humans , Immunoglobulin Heavy Chains , Genetics , Immunoglobulin kappa-Chains , Genetics , Peptide Library , Urinary Bladder Neoplasms , Genetics , Allergy and Immunology
13.
Article in English | WPRIM | ID: wpr-33794

ABSTRACT

Light chain deposition disease (LCDD) is a rare disorder associated with a clonal proliferation of plasma cells, which synthesize abnormal monoclonal immunoglobulin light chains. LCDD is characterized by systemic deposition of light chains in various organs, with the kidneys being most commonly affected. There have been few reports of isolated LCDD. We report a rare case of LCDD limited to a duodenal polyp. A 63-yr-old man visited our hospital for health screening without symptoms in 2009. On gastrofiberscopy, a duodenal polyp was observed. The biopsy showed diffuse infiltration by atypical plasma cells, which were positive for kappa-type light chains by immunohistochemistry. While the patient refused further management, we could find no evidence of recurrence until 2 yr after the initial diagnosis. It has been reported that isolated LCDD has relatively good prognosis compared to systemic LCDD. However, treatment for this disease has not been established yet.


Subject(s)
Duodenum/pathology , Endoscopy, Gastrointestinal , Humans , Immunoglobulin kappa-Chains/immunology , Immunohistochemistry , Intestinal Mucosa/pathology , Male , Middle Aged , Paraproteinemias/diagnosis , Tomography, X-Ray Computed
14.
Article in Chinese | WPRIM | ID: wpr-315529

ABSTRACT

<p><b>OBJECTIVE</b>To construct a personalized full-length fully human antibody mammalian display library for children with systemic lupus erythematosus (SLE).</p><p><b>METHODS</b>The total RNA was isolated from the PBMCs of SLE children. The heavy chain variable region and kappa light chain (VH and LCκ) of the antibody genes were amplified by RT-PCR and inserted into the pDGB-HC-TM vector separately to construct the heavy chain and light chain libraries. The library DNAs were transfected into 293T cells and the expression of full-length fully human antibody on the surface of 293T cells was analyzed by flow cytometry.</p><p><b>RESULTS</b>Using 0.8 µg total RNA as the template, the VH and LCκ were amplified and the full-length fully human antibody mammalian display library was constructed. The VH and LCκ gene libraries had a size of 9.4×10(4) and 8.4×10(4), respectively. Sequence analysis of 10 clones randomly selected from the VH and LCκ gene libraries each showed that 8 heavy chain clones and 7 light chain clones contained correct open reading frames, and flow cytometry demonstrated that all the 15 clones express full-length antibodies on 293T cell surfaces. 293T cells co-transfected with the VH and LCκ gene libraries expressed the full-length antibodies on the cell surface.</p><p><b>CONCLUSION</b>The personalized full-length fully human antibody library for SLE children constructed allows display of the full-length antibodies on mammalian cell surfaces, thus providing a valuable platform for analyzing the autoantibodies, their etiological role, and their clinical implications in SLE.</p>


Subject(s)
Amino Acid Sequence , Child , Gene Library , Genetic Vectors , Humans , Immunoglobulin Heavy Chains , Genetics , Immunoglobulin kappa-Chains , Genetics , Lupus Erythematosus, Systemic , Genetics , Allergy and Immunology , Membrane Proteins , Genetics
15.
Article in English | WPRIM | ID: wpr-315457

ABSTRACT

<p><b>OBJECTIVE</b>To investigate the role of bone morphogenetic protein 4 (BMP4) in culturing induced pluripotent stem cells (iPSCs) and the related signal pathways.</p><p><b>METHODS</b>We amplified the mature peptide of BMP4 from the placenta through RT-PCR, and IgK secretion peptide was ligated to the N-terminal of BMP4 mature peptide. The recombinant plasmid pPYCAG-IgK-BMP4 was transfected into 293T cells and screened with puromycin, and the positive clones for expressing BMP4 were verified by cell immunofluorescence and Western blotting. To test the bioactivity of BMP4, iPSCs were cultured in the medium supplemented with leukemia inhibitory factor (LIF) plus the supernatant containing BMP4, and the cell phenotype, cell differentiation capacity into lineages of the 3 germ layers and expression levels of pluripotency-associated genes were investigated.</p><p><b>RESULTS</b>Smad1 was phosphorylated by BMP4 from the culture medium. iPSCs cultured in the medium supplemented with LIF plus the supernatant containing BMP4 for 3 passages maintained the phenotype of stem cells with the expression levels of pluripotency-associated genes not affected. These iPSCs also maintained the capacity to differentiate into cell lineages of the 3 germ layers.</p><p><b>CONCLUSION</b>BMP4 can be efficiently expressed in mammalian cells to maintain the multipotent differentiation capacity of the iPSCs in in vitro culture.</p>


Subject(s)
Animals , Bone Morphogenetic Protein 4 , Genetics , Cell Differentiation , Genetics , Cells, Cultured , Female , Gene Expression , HEK293 Cells , Humans , Immunoglobulin kappa-Chains , Genetics , Induced Pluripotent Stem Cells , Cell Biology , Mice
16.
Article in English | WPRIM | ID: wpr-164047

ABSTRACT

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.


Subject(s)
Adenocarcinoma/complications , Antineoplastic Agents/therapeutic use , Bone Marrow Cells/metabolism , Combined Modality Therapy , Humans , Immunoelectrophoresis , Immunoglobulin kappa-Chains/blood , Immunoglobulin lambda-Chains/blood , Male , Middle Aged , Multiple Myeloma/complications , Neoplasm Staging , Positron-Emission Tomography , Prostatic Neoplasms/complications , Spine/pathology , Syndecan-1/metabolism , Tomography, X-Ray Computed
17.
Article in Chinese | WPRIM | ID: wpr-235121

ABSTRACT

<p><b>OBJECTIVE</b>To construct a rheumatoid arthritis-specific full-length fully human mammalian display antibody libraries.</p><p><b>METHODS</b>Peripheral blood lymphocytes were isolated from patients with rheumatoid arthritis. The repertoires of kappa light chain (LCκ) and heavy chain variable region (VH) of the antibodies were amplified by RT-PCR. The amplified LCκ and VH genes were inserted into the vector pDGB-HC-TM separately, and the ligated libraries were transformed into competent E.coli TOPO-10 strain to construct the rheumatoid arthritis-specific antibody heavy and light chain libraries. 293T cells were co-transfected with the libraries and the full-length fully human antibody expressed on the surface of 293T cells were analyzed by flow cytometry.</p><p><b>RESULTS</b>The libraries of rheumatoid arthritis-specific antibody LCκ and heavy chain (IgG1) were constructed. The expression of full-length fully human antibody on the surface of 293T cells was confirmed by flow cytometry. With the rates of correct LCκ and heavy chain sequence insertion reaching 80% and 60%, respectively, as shown by DNA sequence analysis of the randomly selected clones, the libraries showed an expressible combinatory diversity of 6.13×10(10).</p><p><b>CONCLUSION</b>The constructed libraries provide a useful platform for screening rheumatoid arthritis-specific antibodies.</p>


Subject(s)
Amino Acid Sequence , Animals , Antibodies , Genetics , Allergy and Immunology , Antibody Specificity , Arthritis, Rheumatoid , Allergy and Immunology , Cell Surface Display Techniques , Cloning, Molecular , Escherichia coli , Genetics , Metabolism , Genetic Vectors , Genetics , HEK293 Cells , Humans , Immunoglobulin G , Genetics , Immunoglobulin Heavy Chains , Genetics , Immunoglobulin kappa-Chains , Genetics , Lymphocytes , Allergy and Immunology , Metabolism , Molecular Sequence Data , Peptide Library , Recombinant Proteins , Genetics , Allergy and Immunology , Transfection
18.
Article in English | IMSEAR | ID: sea-139903

ABSTRACT

Plasma cell granuloma is a rare reactive lesion composed of polyclonal plasma cells. It manifests primarily in the lungs, but may occur in various other anatomic locations like the oral cavity. Intraoral plasma cell granulomas involving the tongue, lip, oral mucosa and gingiva have been reported in the past. This case presents a 54-year-old female with chronic periodontitis and mandibular anterior gingival overgrowth treated by Phase I therapy (scaling and root planing) and excisional biopsy. Histological examination revealed inflammatory cell infiltrate containing sheets of plasma cells. Immunohistochemistry for kappa and lambda light chains showed a polyclonal staining pattern confirming a diagnosis of plasma cell granuloma. This case highlights the need to biopsy for unusual lesions to rule out potential neoplasms.


Subject(s)
Chronic Periodontitis/diagnosis , Diagnosis, Differential , Female , Gingival Diseases/diagnosis , Gingival Diseases/pathology , Gingival Overgrowth/diagnosis , Granuloma, Plasma Cell/diagnosis , Granuloma, Plasma Cell/pathology , Humans , Immunoglobulin kappa-Chains/analysis , Immunoglobulin lambda-Chains/analysis , Middle Aged
19.
Chinese Journal of Cancer ; (12): 1029-1034, 2010.
Article in English | WPRIM | ID: wpr-296320

ABSTRACT

Extramedullary plasmacytoma of the larynx is rare, especially when coexisted with squamous cell carcinoma in situ. We report a 56-year-old woman with hoarseness for 6 months and dysphonia for a week. Fiberoptic laryngoscopic examination showed a red, smooth-surface swelling in the submucous region of the left ventricle and ventricular band of the larynx. The patient underwent vertical laryngectomy and modified left neck dissection. Postoperative pathologic examination revealed coexisting plasmacytoma and carcinoma in situ. Bone marrow biopsy and systemic radiogram showed no positive findings. The hepatic and renal functions were normal. Monoclonal immunoglobulin light chain of type kappa was detected in urine. Hence, a laryngeal extramedullary plasmacytoma with carcinoma in situ was diagnosed. No recurrence or progression was observed during a 2-year follow-up. Here, we discussed the risk factors, diagnosis, and therapy for this rare disease.


Subject(s)
ADP-ribosyl Cyclase 1 , Metabolism , Carcinoma in Situ , Diagnosis , Diagnostic Imaging , Metabolism , Pathology , General Surgery , Carcinoma, Squamous Cell , Diagnosis , Diagnostic Imaging , Metabolism , Pathology , General Surgery , Female , Follow-Up Studies , Humans , Immunoglobulin A , Metabolism , Immunoglobulin kappa-Chains , Metabolism , Laryngeal Neoplasms , Diagnosis , Diagnostic Imaging , Metabolism , Pathology , General Surgery , Laryngectomy , Laryngoscopy , Middle Aged , Mucin-1 , Metabolism , Neck Dissection , Plasmacytoma , Diagnosis , Diagnostic Imaging , Metabolism , Pathology , General Surgery , Syndecan-1 , Metabolism , Tomography, X-Ray Computed
20.
Korean Journal of Dermatology ; : 1114-1117, 2010.
Article in Korean | WPRIM | ID: wpr-23198

ABSTRACT

Primary cutaneous plasmacytoma is a rare type of cutaneous B-cell lymphoma that arises primarily in the skin, and this is derived from clonally expanded plasma cells with various degrees of maturation and atypia. A 72-year-old man had an asymptomatic, solitary reddish to violaceous nodule on the back for about 4 months. The histologic finding of the skin biopsy specimen demonstrated an infiltration of variably matured plasma cells in the dermis, and these cells showed a monotypic expression of immunoglobulin kappa chains on immunohistochemical staining. Staging investigations excluded any extracutaneous manifestations of the disease. Only a few cases of primary cutaneous plasmacytoma have been published in the Korean dermatologic literature. Herein, we report on an interesting case of primary cutaneous plasmacytoma with monoclonality of kappa chains.


Subject(s)
Aged , Biopsy , Dermis , Humans , Immunoglobulin kappa-Chains , Lymphoma, B-Cell , Plasma Cells , Plasmacytoma , Skin
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