Immunoglobulin Kappa Research Articles

Population matched (pm) germline allelic variants of immunoglobulin (IG) loci: Relevance in infectious diseases and vaccination studies in human populations

Immunoglobulin (IG) loci harbor inter-individual allelic variants in many different germline IG variable, diversity and joining genes of the IG heavy (IGH), kappa (IGK) and lambda (IGL) loci, which together form the genetic basis of the highly diverse antigen-specific B-cell receptors. These allelic variants can be shared between or be specific to human populations. The current immunogenetics resources gather the germline alleles, however, lack the population specificity of the alleles which poses limitations for disease-association studies related to immune responses in different human populations. Therefore, we systematically identified germline alleles from 26 different human populations around the world, profiled by “1000 Genomes” data. We identified 409 IGHV, 179 IGKV, and 199 IGLV germline alleles supported by at least seven haplotypes. The diversity of germline alleles is the highest in Africans. Remarkably, the variants in the identified novel alleles show strikingly conserved patterns, the same as found in other IG databases, suggesting over-time evolutionary selection processes. We could relate the genetic variants to population-specific immune responses, e.g. IGHV1-69 for flu in Africans. The population matched IG (pmIG) resource will enhance our understanding of the SHM-related B-cell receptor selection processes in (infectious) diseases and vaccination within and between different human populations.

A proteomic analysis of urine biomarkers in autism spectrum disorder

Autism spectrum disorder (ASD) is a neurodevelopmental condition characterized by early-onset social-communication challenges, restricted and repetitive behaviors, or unusual sensory-motor behaviors. A lack of specific biomarkers hinders the early diagnosis and treatment of this disease in many children. This study analyzes and validates potential urinary biomarkers using mass spectrometry proteomics. Global proteomics profiles of urine from 19 ASD patients and 19 healthy control subjects were compared to identify significantly changed proteins. These proteins were validated with targeted proteomics using parallel reaction monitoring (PRM) in an independent validation set consisting of samples from 40 ASD patients and 38 healthy controls. A total of 34 significantly changed proteins were found in the discovery set, among which seven proteins were identified as potential biomarkers for ASD through PRM assays in the validation set. Of these seven proteins, immunoglobulin kappa variable 4–1, immunoglobulin kappa variable 3–20, and immunoglobulin lambda variable 1–51 were up-regulated, while ATP synthase F1 subunit alpha, 10 kDa heat shock protein, apolipoprotein C-III, and arylsulfatase F were down-regulated. Six of these seven proteins support previous findings that ASD is accompanied by altered immune response and lipid metabolism, as well as mitochondrial dysfunction. This study lays the groundwork for additional research using biomarkers to clinically diagnose ASD. The proteomics and PRM raw data of this study have been deposited under the accession number IPX0002592000 at iProX. SignificanceThis study identified 34 proteins in urine of ASD patients that were significantly changed from the urinary proteins of healthy subjects using LC-MS/MS-based proteomics in a discovery set. Seven of these proteins were validated by PRM analysis in an independent validation set. This report represents the first description of combined label-free quantitative proteomics and PRM analysis of targeted proteins for discovery of ASD urinary biomarkers. The results will be helpful for early diagnosis and can provide additional insight into the molecular mechanisms of ASD.

Issue Highlights — May 2021

Issue Highlights — May 2021

Elevated RBP-Jκ and CXCL11 Expression in Colon Cancer is Associated with an Unfavorable Clinical Outcome.

IntroductionThis study aims at exploring the expression and significance of recombination signal-binding protein for immunoglobulin kappa J region (RBP-Jκ) and C-X-C motif chemokine 11 (CXCL11) in human colon cancer tissues.MethodsThe RBP-Jκ and CXCL11 expression levels were assessed by immunohistochemistry and quantitative real-time polymerase chain reaction (qRT-PCR) in patients with colon cancer, and their prognostic significance was evaluated.ResultsThrough analyzing 342 samples of colon cancer patients treated at our institution, increased expression of RBP-Jκ and CXCL11 was found in human colon cancer specimens compared with matched paratumorous normal specimens (P<0.001). A positive correlation was found between RBP-Jκ expression and CXCL11 expression (P<0.001). High RBP-Jκ expression was significantly associated with poorly differentiated tumors (P=0.005), invasion beyond propria muscularis (P=0.025), lymph node metastases (P=0.005), distant metastasis (P<0.001), advanced tumor-node-metastasis (TNM) stage (P=0.004), and a shorter overall survival (P<0.001). An increase in CXCL11 protein expression was associated with poorly differentiated tumors (P=0.015), invasion beyond propria muscularis (P=0.029), lymph node metastases (P=0.031), distant metastasis (P=0.045), advanced TNM stage (P=0.026), and a shorter overall survival of patients (P<0.001). In multivariate Cox regression analysis, RBP-Jκ protein expression (P=0.036), CXCL11 protein expression (P=0.001), differentiation (P<0.001), depth of invasion (P=0.009), distant metastasis (P<0.001), and TNM stage (P<0.001) were independent prognostic indicators of colon cancer.ConclusionHigh expression of RBP-Jκ is closely associated with high CXCL11 expression, which represents a risk factor for the poor overall survival of colon cancer patients.

Prophylactic administration of fingolimod (FTY720) ameliorated experimental autoimmune myasthenia gravis by reducing the number of dendritic cells, follicular T helper cells and antibody-secreting cells

Fingolimod (FTY720), a sphingosine 1-phosphate (S1P) receptor antagonist, possesses potent immunomodulatory activity via lymphocyte homing. The effects of FTY720 have been widely studied in various T-cell-mediated autoimmune diseases, while the immunomodulatory effects on experimental autoimmune myasthenia gravis (EAMG), a typical disease model for antibody-mediated autoimmunity, remain elusive. In the present study, FTY720 was administered to EAMG rats as prophylaxis. The clinical scores were recorded every other day, and serum antibodies at different time points were measured by enzyme-linked immunosorbent assay (ELISA). The immune cell subsets in the spleen, bone marrow, circulation, and thymus were determined by flow cytometry. The prophylactic administration alleviated EAMG symptoms by reducing the level of serum antibodies IgG and its isotype IgG2b on days 30 and 46 post immunization, as well as IgG and Ig kappa antibody-secreting cells in the spleen and bone marrow. The mitigated humoral immune response can be attributed to the decreased dendritic cells, follicular T help cells (Tfh) and Tfh subsets (Tfh1, Tfh2, and Tfh17), and T helper cell subsets (Th1, Th2, and Th17) in the spleen. The promotion of lymphocyte homing and inhibition of thymocyte egress contribute to the effects of FTY720 on these effector T cell subsets. Overall, the prophylactic administration of FTY720 ameliorated EAMG partially by regulating humoral immune response，suggesting that FTY720 could be part of a pharmacological strategy for managing myasthenia gravis.

NOTCH signaling pathway is required for bovine early embryonic development†.

The NOTCH signaling pathway plays an important role in regulating various biological processes, including lineage specification and apoptosis. Multiple components of the NOTCH pathway have been identified in mammalian preimplantation embryos. However, the precise role of the NOTCH pathway in early embryonic development is poorly understood, especially in large animals. Here, we show that the expression of genes encoding key transcripts of the NOTCH pathway is dynamic throughout early embryonic development. We also confirm the presence of active NOTCH1 and RBPJ. By using pharmacological and RNA interference tools, we demonstrate that the NOTCH pathway is required for the proper development of bovine early embryos. This functional consequence could be partly attributed to the major transcriptional mediator, Recombination Signal Binding Protein For Immunoglobulin Kappa J Region (RBPJ), whose deficiency also compromised the embryo quality. Indeed, both NOTCH1 and RBPJ knockdown cause a significant increase of histone H3 serine 10 phosphorylation (pH3S10, a mitosis marker) positive blastomeres, suggesting a cell cycle arrest at mitosis. Importantly, RNA sequencing analyses reveal that either NOTCH1 or RBPJ depletion triggers a reduction in H1FOO that encodes the oocyte-specific linker histone H1 variant. Interestingly, depleting H1FOO results in detrimental effects on the developmental competence of early embryos, similar with NOTCH1 inhibition. Overall, our results reveal a crucial role for NOTCH pathway in regulating bovine preimplantation development, likely by controlling cell proliferation and maintaining H1FOO expression.

Predictive Markers of Missed Miscarriage

The aim of this study was to find useful the serological markers for missed miscarriage (MM) in order to predict the outcome of pregnancy. The study included 141 pregnant women aged between 18 and 45 years at gestational age under 11 weeks. All women were divided into 3 groups. Group 1 included 68 women with MM; Group 2 included 43 women with spontaneous miscarriage; Group 3 included 30 pregnant women without pathology. Proteomic analysis of the blood serum was performed using liquid chromatography-mass spectrometry. The results of our study show that immunoglobulin kappa variable 3-15 (KV315) can be considered as the most promising serologic marker for MM in early gestation. The potential role of KV315 as the serological marker is very important for predicting the course of pregnancy.

A154 PROTEOMIC IDENTIFICATION OF INFLAMMATORY AND FIBROSTENOTIC BLOOD SERUM BIOMARKERS IN CROHN’S DISEASE

Abstract Background Crohn’s disease (CD) is an incurable relapsing-remitting inflammatory bowel disease (IBD) where patients may experience bowel damage with symptoms such as abdominal pain, chronic diarrhea, extraintestinal manifestations and life-long disability. CD is heterogeneous with three distinct phenotypes including a stricturing phenotype marked by intestinal fibrosis. This fibrotic morphology is generally more unresponsive to drug treatment; delaying a patient’s remission, control of the disease and often requiring surgical intervention. Thus, early and accurate identification of fibrostenosis in CD is important to optimize patient treatment and predict response to therapy. Aims The aim of our study is to distinguish inflammatory and intestinal fibrostenosis in CD patients using serum protein biomarkers. Methods Blood sera from 17 inflammatory and 17 fibrostenotic CD patients were collected. The phenotypic classification was confirmed by intestinal ultrasound and endoscopy. Samples were subjected to Shotgun Proteomics, an unbiased proteomics approach that allows for relative protein quantification. Proteins from each condition were isotopically labelled with formaldehyde (light +28 Da and heavy +34 Da), pooled and digested with trypsin. Following liquid chromatography and tandem mass spectrometry, peptides were then identified by MaxQuant software with a false discovery rate of 1%. Excel (Microsoft), Prism (Graphpad) and Metascape software were used for data filtering and analysis. Results Proteomic processing allowed for the identification of novel protein biomarkers in the inflammatory and stricture CD phenotypes. Inflammation was correlated with activation of the complement pathway and fatty acid metabolism; marked by increased levels of immunoglobulin gamma 4 chain (IGHG4), mitochondrial creatinine kinase (CKMT1), apolipoprotein A (LPA) and glutathione peroxidase 3 (GPX3) proteins. Fibrostenosis showed no distinct metabolic pathway, but an elevated expression of SWI/SNF-related matric associated actin (SMARCA5), haptoglobin related protein (HPR), immunoglobulin kappa and immunoglobulin heavy constant proteins. Conclusions Our data indicate that inflammation and strictures in CD may be driven by distinct signaling pathways. We identified specific protein signatures for the two phenotypes, which may aid in predicting those who are at risk of developing strictures and in the development of phenotype-specific treatment for CD patients. Future validation of these proteins will be performed to assess this unique protein profile. Funding Agencies McCaig Institute for Bone and Joint Health

Isotypes of autoantibodies against novel differential 4-hydroxy-2-nonenal-modified peptide adducts in serum is associated with rheumatoid arthritis in Taiwanese women

BackgroundRheumatoid arthritis (RA) is an autoimmune disorder with systemic inflammation and may be induced by oxidative stress that affects an inflamed joint. Our objectives were to examine isotypes of autoantibodies against 4-hydroxy-2-nonenal (HNE) modifications in RA and associate them with increased levels of autoantibodies in RA patients.MethodsSerum samples from 155 female patients [60 with RA, 35 with osteoarthritis (OA), and 60 healthy controls (HCs)] were obtained. Four novel differential HNE-modified peptide adducts, complement factor H (CFAH)1211–1230, haptoglobin (HPT)78–108, immunoglobulin (Ig) kappa chain C region (IGKC)2–19, and prothrombin (THRB)328–345, were re-analyzed using tandem mass spectrometric (MS/MS) spectra (ProteomeXchange: PXD004546) from RA patients vs. HCs. Further, we determined serum protein levels of CFAH, HPT, IGKC and THRB, HNE-protein adducts, and autoantibodies against unmodified and HNE-modified peptides. Significant correlations and odds ratios (ORs) were calculated.ResultsLevels of HPT in RA patients were greatly higher than the levels in HCs. Levels of HNE-protein adducts and autoantibodies in RA patients were significantly greater than those of HCs. IgM anti-HPT78−108 HNE, IgM anti-IGKC2−19, and IgM anti-IGKC2−19 HNE may be considered as diagnostic biomarkers for RA. Importantly, elevated levels of IgM anti-HPT78−108 HNE, IgM anti-IGKC2−19, and IgG anti-THRB328−345 were positively correlated with the disease activity score in 28 joints for C-reactive protein (DAS28-CRP). Further, the ORs of RA development through IgM anti-HPT78−108 HNE (OR 5.235, p < 0.001), IgM anti-IGKC2−19 (OR 12.655, p < 0.001), and IgG anti-THRB328−345 (OR 5.761, p < 0.001) showed an increased risk. Lastly, we incorporated three machine learning models to differentiate RA from HC and OA, and performed feature selection to determine discriminative features. Experimental results showed that our proposed method achieved an area under the receiver operating characteristic curve of 0.92, which demonstrated that our selected autoantibodies combined with machine learning can efficiently detect RA.ConclusionsThis study discovered that some IgG- and IgM-NAAs and anti-HNE M-NAAs may be correlated with inflammation and disease activity in RA. Moreover, our findings suggested that IgM anti-HPT78−108 HNE, IgM anti-IGKC2−19, and IgG anti-THRB328−345 may play heavy roles in RA development.

LncRNA FTX Promotes Colorectal Cancer Cells Migration and Invasion by miRNA-590-5p/RBPJ Axis.

Colorectal cancer (CRC) is one of the most lethal human cancers all over the world. Moreover, it ranks fourth for cancer-related deaths among males. Although many efforts have been made to cure CRC, the effect remains limited. It has been reported that lncRNA five prime to Xist (FTX) was upregulated in CRC. However, the mechanism by which lncRNA FTX regulates the progression of CRC remains largely unknown. In this study,qRT-PCR was performed to detect the expressionof FTX, miR-590-5p andRecombination signal binding protein for immunoglobulin kappa J region (RBPJ) in CRC tissues or cells. Protein expression in cells was measured by western blot. MTT assay was used to test the cell viability. Moreover, transwell was performed to examine the cell migration and invasion. Luciferase report assay was performed to verify the relation between miR-590-5p and FTX or RBPJ. It was found thatFTX was upregulated in CRC tissues and cells. Knockdown of FTX or overexpression of miR-590-5p can inhibit the proliferation, migration, and invasion of CRC cells. Besides, silencing of FTX could inhibit the expression of migration and invasion-related proteins in CRC cells. Meanwhile, miR-590-5p was the target of FTX, and RBPJ was the direct target of miR-590-5p. Inhibition of miR-590-5p could reverse the inhibitory effect of FTX on the progression of CRC. These findings suggested that knockdown of FTX could inhibit the tumorigenesis of CRC in vitro, which may serve as a potential novel strategy for treatment of CRC.

Heart damage in the combined types of systemic amyloidosis

To identify and determine the clinical and morphological characteristics of combined types of SA in patients with biventricular chronic heart failure (CHF). Eighty autopsy protocols for biventricular CHF deaths were retrospectively analyzed. Immunohistochemistry and confocal laser scanning microscopy (CLCM) with antibodies to amyloid A (AA), serum amyloid-P (SAP), prealbumin, and immunoglobulin kappa (κ) and lambda (λ) light chains were performed. The myocardium showed a combination of different types of SA in 6 (7.5%) cases, including Alλ-CA+ATTR-CA, ALκ-CA+ATTR-CA, and AA-CA+ATTR-CA in 4, 1, and 1 cases, respectively. Macroscopically, the heart mass averaged 470±20 g; the thickness of the left and right ventricular myocardium was 1.5±0.2 and 0.4±0.1 cm, respectively; the interventricular septum averaged 1.2±0.2 cm; and the cardiac index was 0.008. The myocardium was dense, dark red with diffuse layers of whitish dense fibrous connective tissue; the heart cavities were enlarged. Microscopically, in 25% of cases, all heart parts had ALλ-CA that was characterized by massive amyloid deposits localized predominantly in the intramyocardial vessel wall, intermuscular connective tissue, and perivascularly. The myocardium also displayed small amyloid deposits of ALλ-CA and ATTR-CA in the intermuscular connective tissue and intramyocardial vessel wall. Amyloid deposits were located in different parts of the myocardium; there were also areas of co-localization of ALλ-CA+ATTR-CA. The combined types of SA occurred under the guise of coronary heart disease and the dilated cardiomyopathy phenotype. The combined amyloid AL-CA and ATTR-CA was generally localized in the interstitium and myocardial vessels. There were also small areas of co-localization of amyloid deposits, which were found mainly in the intramyocardial vessels.

Autopsy findings in a patient with primary systemic AL (kappa light chain) amyloidosis.

ABSTRACTFirst described by Rokitansky in 1842, and further characterized by Virchow in 1854, amyloidosis is a disorder caused by amyloid deposition, a fibrillary insoluble protein. The clinical spectrum of amyloidosis is broad, as the amyloid deposition may virtually occur in all tissues. Herein, we report the case of a 66-year-old man with a long-lasting emaciating disease, diagnosed, at autopsy, with primary systemic amyloidosis. Amyloid protein deposition was found in many tissues and organs. The involvement of the vessels’ wall rendered ischemic injury most prominent in the intestinal loops causing mesenteric ischemia. Despite the thorough organic involvement, the immediate cause of death was aspiration bronchopneumonia. Massive amyloid deposition was found in virtually all major organs, such as the heart, liver, kidneys, spleen, pancreas, adrenals, prostate, skin, and thyroid: the latter, a complication of the amyloidosis known as amyloid goiter. Post-mortem review of the deceased’s laboratory workup showed a slightly abnormal kappa:lambda ratio in the blood; however, no clonal lymphoplasmacytic disorder was confirmed in the bone marrow and other lymphoreticular system organs either by the microscopic examination and immunohistochemical staining. Laser-capture microdissection and tandem mass spectrometry of the splenic tissue detected a peptide profile consistent with an immunoglobulin Kappa light chain. The presence of amyloid purpura favors the diagnosis of primary systemic amyloidosis.

Association of a Functional Single Nucleotide Polymorphism (rs874040) in the RBPJ Gene with Susceptibility to Rheumatoid Arthritis in Iranian Population.

Background:Rheumatoid Arthritis (RA) is a progressive, heterogeneous, and common multifactorial autoimmune disease. Several Genome-Wide Association Studies (GWASs) have revealed more than 100 risk loci for RA. One of these loci is a functional single nucleotide polymorphism (rs874040; G>C) near the recombination signal-binding protein for the immunoglobulin kappa J region (RBPJ) gene. RBPJ can convert into a transcriptional activator upon activation of the canonical Notch pathway. Notch signaling has recently emerged as an important regulator of immune responses in inflammation and autoimmune diseases. In the present study, the possible association between SNP rs874040 (G>C) upstream of the RBPJ gene with RA risk was assessed in Iranian population.Methods:A case-control study including 60 RA patients and 44 control subjects was conducted to estimate rs874040 genotypes using real-time polymerase chain reaction High Resolution Melting (HRM) method.Results:Logistic regression analysis indicated that homozygous CC and heterozygous GC genotypes increase the risk of RA compared with GG genotype (CC vs. GG; OR=11.36; 95% CI [3.93–33.33] and CG vs. GG; OR=3.78; 95% CI [1.30–10. 98]). Besides, subjects with C allele were more frequently affected with RA than subjects with G allele (OR=10.42; 95% CI [5.21–20.83]). Furthermore, in the patient group, a significant correlation was found between C-reactive protein concentrations and rs874040 polymorphism (p<0.05).Conclusion:Our findings propose a substantial correlation between rs874040 polymorphism and RA risk in Iranian population.

Isotyping and Semi-Quantitation of Monkey Anti-Drug Antibodies by Immunocapture Liquid Chromatography-Mass Spectrometry

There is an urgent demand to develop new technologies to characterize immunogenicity to biotherapeutics. Here, we developed an immunocapture LC-MS assay to isotype and semi-quantify monkey anti-drug antibodies (ADAs) to fully human monoclonal antibody (mAb) drugs. ADAs were isolated from serum samples using an immunocapture step with the Fab of the full-length mAb cross-linked to magnetic beads to minimize matrix interference. A positive monoclonal antibody control against the human immunoglobulin kappa light chain was used as a calibration standard for ADA quantitation. The final LC-MS method contains 17 multiple reaction monitoring (MRM) transitions and an optimized 15-min LC method. The results suggested that IgG1 was the most abundant isotype in ADA-positive samples. IgG2 and IgG4 were identified at lower levels, whereas IgG3 and IgA levels were only observed at very minor levels. In addition, levels of total ADA measured by the LC-MS assay were comparable to results obtained using a traditional ligand binding assay (LBA). The LC-MS ADA assay enabled rapid immunogenicity assessment with additional isotype information that LBAs cannot provide.

Prognostic Impact of Immunoglobulin Kappa C in Breast Cancer Patients Treated with Adjuvant Chemotherapy

Introduction: Immunoglobulin κC (IGKC)-positive tumor-infiltrating plasma cells are associated with better prognosis in node-negative breast cancer patients without adjuvant systemic therapy. In the present study we evaluated the prognostic significance of IGKC in breast cancer patients treated with adjuvant chemotherapy ± tamoxifen. Methods: IGKC expression was immunohistochemically analyzed in 193 breast cancer patients who were treated with adjuvant chemotherapy, either with cyclophosphamide, methotrexate, and 5-fluorouracil (CMF) or 5-fluorouracil, epirubicin, and cyclophosphamide (FEC) between 1993 and 2001 with a median follow-up of 11 years. The prognostic impact of IGKC expression was evaluated by Kaplan-Meier survival analyses as well as uni- and multivariate Cox regression. An interaction term was used to investigate a possible association between tamoxifen treatment and prognostic effect of IGKC expression. Results: Kaplan-Meier analyses identified IGKC as a prognostic marker for metastasis-free survival (MFS): higher IGKC expression was associated with a better outcome (p = 0.02, log rank). Results of univariate Cox regression confirmed the prognostic impact of IGKC expression: patients with a strong IGKC expression had a longer MFS (hazard ratio [HR] 1.931; 95% confidence interval [CI] 1.087–3.431; p = 0.025). Multivariate Cox regression analysis showed the independent prognostic significance of IGKC expression (HR 2.070; 95% CI 1.088–3.938; p = 0.027). The interaction term confirmed a significant interaction between tamoxifen treatment and the prognostic impact of IGKC expression (p_interaction = 0.04). Conclusion: IGKC expression had an independent prognostic impact in early breast cancer patients who received adjuvant chemotherapy. There was a significant interaction with the use of tamoxifen.

Clinical and biological features of B-cell neoplasms with CDK6 translocations: an association with a subgroup of splenic marginal zone lymphomas displaying frequent CD5 expression, prolymphocytic cells, and TP53 abnormalities.

A translocation involving the cyclin-dependent kinase 6 (CDK6) gene [t(CDK6)] is a rare but recurrent abnormality in B-cell neoplasms. To further characterise this aberration, we studied 57 cases; the largest series reported to date. Fluorescence in situ hybridisation analysis confirmed the involvement of CDK6 in all cases, including t(2;7)(p11;q21) immunoglobulin kappa locus (IGK)/CDK6 (n=51), t(7;14)(q21;q32) CDK6/immunoglobulin heavy locus (IGH) (n=2) and the previously undescribed t(7;14)(q21;q11) CDK6/T-cell receptor alpha locus (TRA)/T-cell receptor delta locus (TRD) (n=4). In total, 10 patients were diagnosed with chronic lymphocytic leukaemia, monoclonal B-cell lymphocytosis or small lymphocytic lymphoma, and 47 had small B-cell lymphoma (SmBL) including 36 cases of marginal zone lymphoma (MZL; 34 splenic MZLs, one nodal MZL and one bronchus-associated lymphoid tissue lymphoma). In all, 18 of the 26 cytologically reviewed cases of MZL (69%) had an atypical aspect with prolymphocytic cells. Among the 47 patients with MZL/SmBL, CD5 expression was found in 26 (55%) and the tumour protein p53 (TP53) deletion in 22 (47%). The TP53 gene was mutated in 10/30 (33%); the 7q deletion was detected in only one case, and no Notch receptor 2 (NOTCH2) mutations were found. Immunoglobulin heavy-chain variable-region (IGHV) locus sequencing revealed that none harboured an IGHV1-02*04 gene. Overall survival was 82% at 10years and not influenced by TP53 aberration. Our present findings suggest that most t(CDK6)+ neoplasms correspond to a particular subgroup of indolent marginal zone B-cell lymphomas with distinctive features.

Asymptomatic pink nodules on a 28-year-old patient

Asymptomatic pink nodules on a 28-year-old patient

Clinicopathological analysis of mucosa associated lymphoid tissue lymphoma secondary to Sjögren' s syndrome in salivary gland

OBJECTIVE To analyze the clinicopathological characteristics of mucosa associated lymphoid tissue (MALT) lymphoma secondary to Sjogren' s syndrome (SS) (SS-MALT lymphoma) in salivary gland and to explore the value of the combined application of histopathological morphology, protein expression and molecular phenotype in pathological diagnosis and prognostic evaluation of SS-MALT lymphoma. METHODS Sixteen patients with SS-MALT lymphoma were collected from 260 patients who were diagnosed with SS in Peking University School and Hospital of Stomatology from January 1997 to December 2016. Twelve patients with non-MALT lymphoma secondary to SS (non-SS-MALT lymphoma) in salivary gland were selected as controls. The clinical data of the patients were retrospectively reviewed and analyzed. All the patients were followed up until December 20, 2019. Hematoxylin-eosin staining, immunohistochemistry, polymerase chain reaction (PCR) and fluorescence in situ hybridization (FISH) techniques were used to observe the histologic characteristics and to detect the manifestations of light chain restrictive expression, immunoglobulin (Ig) gene clonal rearrangement, chromosome translocation and gene abnormality, so as to evaluate their values in pathological diagnosis and prognostic evaluation. RESULTS The malignant transformation rate of SS to MALT lymphoma was about 6.15%, ranged from 3 to 240 months, during which 2 patients died due to high-level deterioration. Microscopically, the acini of the glandular tissue were atrophied and destroyed. The tumor cells dominated by central cell-like lymphocytes grew diffusely, destroying the epithelial islands. All SS-MALT lymphoma cases were positive in CD20 and Pax5. Half of them had the Ki-67 proliferation index of 10% or less, and half greater than 10%. 93.75% cases expressed AE1/AE3 protein, which showed the residual glandular epithelium. All the tumor cells were negative in CD3e, and the plasma cells were detected by CD138 antigen. The light chain restrictive expression of κ and λ was 37.5% in SS-MALT lymphoma group. The positive detection rates of immunoglobulin heavy chain (IgH)-FR1, IgH-FR2, IgH-FR3, immunoglobulin kappa chain (IgK)-A, and IgK-B in SS-MALT lymphoma group were 33.3%, 53.3%, 33.3%, 20.0%, and 26.7%, respectively, and 93.3% when together used with IgH and IgK. The positive rates of the MALT1, IGH and BCL6 genes with dual color break-apart probes were 36.4%, 27.3% and 27.3%, and the detection rate of chromosome translocation and gene abnormality by applying the three probes was 72.7%. CONCLUSION There are no specific histological characteristics and protein phenotypes in the histologic diagnosis of SS-MALT lymphoma in salivary gland. The combined application of histopathological manifestations, immunohistochemistry, PCR and FISH techniques helps the accurate pathologic diagnosis of the disease. Although SS-MALT lymphoma is considered as an indolent lymphoma with a relatively favorable prognosis, the regular return visit and long-term follow-up should be conducted to detect the clues of recurrence and advanced deterioration.

The application of thermal oscillation method to augment the effectiveness of autologous platelet rich plasma in treating elderly patients with knee osteoarthritis.

Elderly patients with knee osteoarthritis (OA) are often troubled with joint soreness, swelling, weakness, and pain. Knee intra-articular (IA) injection of autologous platelet rich plasma (PRP) is one of the options that can be used in treating knee OA. However, there are doubts on the effect of PRP when used in treating elderly patients with moderate to severe degrees of knee OA. In this study, 47 elderly patients with moderate degree of knee OA combined with supra-patellar bursitis were recruited. Musculoskeletal ultrasound was used for accurate needle placement for the aspiration of bursa synovial fluid SF followed by PRP injections. In group 1, 23 patients received conventional liquid-form IA PRP injections. In group 2, 24 patients received IA injections of thermal oscillation prepared PRP injectants. It was discovered that when the liquid-form PRP was heated up to a temperature of 75°C and simultaneously oscillated under 200 revolutions per minute (rpm) for 15min, the end product became paste-form like and with a viscosity similar to that of an anti-cough syrup. Under incubation, the highest number of platelet-derived growth factor (PDGF) was obtained on the 8th day. At one month after the completion of PRP injections, group 2 patients revealed significant decreases in SF total protein concentrations, SF volumes, and Lequesne index values. Proteins associated with inflammation, such as apolipoprotein A-I, haptoglobin, immunoglobulin kappa chain, transferrin, and matrix metalloproteinase also decreased significantly. Therefore, the thermal oscillation preparation method can augment the effectiveness of autologous PRP in treating elderly patients with moderate knee OA. It may be recommended that the PRP injectant should be prepared first using the thermal oscillation method to increase its viscosity, enabling prolonged release of growth factors once it is injected into the knee joint.

COEXISTENCE OF LYMPHOCYTIC INTERSTITIAL PNEUMONITIS, AMYLOIDOSIS, AND MALT LYMPHOMA ASSOCIATED WITH SJÖGREN’S SYNDROME

SESSION TITLE: Fellows Pulmonary Manifestations of Systemic Disease Posters SESSION TYPE: Fellow Case Report Posters PRESENTED ON: October 18-21, 2020 INTRODUCTION: Various pulmonary abnormalities have been linked with Sjögren’s syndrome. Here we describe a case of diffuse cystic lung disease in a patient with long-standing Sjögren’s syndrome. Pathology revealed three different yet interrelated processes. CASE PRESENTATION: A 60-year-old female never-smoker with long-standing Sjögren’s syndrome underwent computed tomography of the chest to investigate cervical lymphadenopathy noted at physical examination. This identified multiple thin-walled pulmonary cysts in both lungs, together with irregular solid nodules (Figure 1). Her pulmonary function test was normal. Immunologic tests were remarkable for positive ANA, ds-DNA, SS-A, and SS-B antibodies. A thoracoscopic lung biopsy revealed extra-nodal marginal zone lymphoma of mucosa-associated lymphoid tissue (MALT) associated with IgA kappa immunoglobulin deposition, arising in a background of lymphocytic interstitial pneumonia (LIP) (Figure 2-A, 2-B). The patient was referred to Hematology and has remained on annual surveillance with no treatment. Three years later, the patient underwent left apical wedge resection to clarify the nature of an enlarging pulmonary nodule (Figure 1-A). Histopathological examination revealed AL (kappa) amyloid deposition. The patient remained asymptomatic until the last follow up, despite no treatment. DISCUSSION: The case illustrates the coexistence of three different but related pathological processes: LIP, amyloidosis and MALT lymphoma. Autoimmunity is associated with chronic immune system stimulation and Sjögren syndrome is recognized as a systemic autoimmune disorder with an increased risk of lymphoma, usually of B cell origin. Meanwhile, abnormal immunoglobulin can be deposited locally in the tissues and cause amyloid. The prognosis for patients with amyloidosis and LIP varies, the condition resolving in some patients, whereas it progresses to pulmonary fibrosis and respiratory failure in others. A variety of immunosuppressants, including but not limited to corticosteroids, cyclophosphamide, rituximab, methotrexate, and azathioprine have been used. Primary pulmonary MALT lymphoma is a rare entity with no standardized therapy. For localized disease, some reported cases had spontaneous regression. For widespread tumors, systemic chemotherapy has been used . CONCLUSIONS: Cystic lung disease is a rare but notable comorbidity in patients with Sjögren syndrome. It can be associated with different coinciding pathological processes. Reference #1: Bertoni F, Zucca E. Delving deeper into MALT lymphoma biology. J Clin Invest. 2006 Jan; 116(1): 22–6. Reference #2: Gupta N, Vassallo R, Wikenheiser-Brokamp KA, et al. Diffuse cystic lung disease. Part II. Am J Respir Crit Care Med. 2015; 192:17–29 Reference #3: Martínez-Balzano, Carlos D., Sunkaru Touray, and Scott Kopec. “Cystic lung disease among patients with Sjögren syndrome: frequency, natural history, and associated risk factors.” Chest 150.3 (2016): 631-639. DISCLOSURES: No relevant relationships by Matthew Cecchini, source=Web Response No relevant relationships by Heyi Li, source=Web Response Recipient of grant funding relationship with Bristol Myers Squibb Please note: >$100000 Added 03/23/2020 by Robert Vassallo, source=Web Response, value=Grant/Research Support Recipient of grant funding relationship with Sun Pharmaceuticals Please note: >$100000 Added 03/23/2020 by Robert Vassallo, source=Web Response, value=Grant/Research Support Recipient of grant funding relationship with Pfizer Please note: >$100000 Added 03/23/2020 by Robert Vassallo, source=Web Response, value=Grant/Research Support