Plasma Cell Count Research Articles

Akkermansia muciniphila Promotes SIgA Production and Alters the Reactivity Towards Commensal Bacteria in Early-Weaned Piglets

BackgroundSecretory IgA (SIgA) is the first line of defense in protecting the intestinal epithelium against pathogenic bacteria, regulating gut microbiota composition, and maintaining intestinal homeostasis. Early weaning strategies may disrupt SIgA levels in piglet intestines, causing a decline in immune response and early weaning stress. However, the specific microbial mechanisms modulating SIgA in early-weaned piglets are not well understood. ObjectiveWe hypothesized that Akkermansia muciniphila increases intestinal SIgA production in the early-weaned piglets. MethodsFecal SIgA levels, SIgA-coated bacteria abundance, and fecal metagenomes were compared between 6 Huanjiang miniature (HM) and 6 Duroc×Landrace×Yorkshire (DLY) early-weaned piglets to identify bacterial species involved in SIgA modulation. 4 bacterial species were investigated using 5 groups (Control, A. muciniphila, L. amylovorus, L. crispatus, L. acidophilus) of male SPF C57BL/6J mice, weaned 3 weeks post-birth (n=8/group). Subsequently, 10-day-old Landrace×Yorkshire (LY) piglets were randomly assigned to three groups (Control, 109A. muciniphila, 108A. muciniphila) (n=10/group) to evaluate the effect of orally administered A. muciniphila on intestinal SIgA production and microbial composition. ResultsHM early-weaned piglets showed significantly higher SIgA levels (7.59 μg/mg, 95% CI: 3.2, 12, P = 0.002) and SIgA-coated bacteria abundance (8.64%, 95% CI: 3.2, 14, P = 0.014) than DLY piglets. In the mouse model, administration of A. muciniphila significantly increased SIgA levels (3.50 μg/mg, 95% CI: 0.59, 6.4, P = 0.018), SIgA-coated bacteria abundance (9.06%, 95% CI: 4, 14, P =0.018), and IgA+ plasma cell counts (6.1%, 95% CI: 4.3, 8, P = 0.005). In the pig experiments, oral administration of A. muciniphila to LY piglets significantly enhanced intestinal SIgA concentrations (4.22μg/mg, 95% CI: 0.37, 8.5, P = 0.034) and altered the SIgA-coated bacterial landscape. ConclusionEarly intervention with A. muciniphila in nursing piglets can increases intestinal SIgA productionand alter the reactivity towards commensal bacteria upon early weaning.

Paucity of gastrointestinal plasma cells in common variable immunodeficiency.

Common variable immunodeficiency enteropathy (CVID-E) is a noninfectious complication of CVID caused by chronic inflammation of the gastrointestinal (GI) tract. Based on literature, a paucity or lack of plasma cells, although not obligatory for diagnosis, is a pathognomonic feature of CVID and more frequent in CVID-E. However, there is no consensus on standardized histopathological analysis of this feature in biopsies. In this systematic review, we highlight methods of reproducible plasma cell quantification of biopsies in CVID and describe the plasma cell counts and classes as presented in the literature. Reduced plasma cell counts are commonly found over the entire GI tract, except for in the oesophagus. Immunoglobulin A+ (IgA+) plasma cells appear to be the most commonly reduced plasma cell class in CVID, yet there is scarce literature on the predictive value of low IgA+ plasma cell counts in CVID-E. We propose two optimized methodologies of quantification using a cut-of value of <10 plasma cells per HPF at 40× magnification, or a proportion of ≥1-5% of total mononuclear cells, recorded over ≥3 sections, and in ≥2 biopsies, as the most conservative agreeable definitions for a paucity of plasma cells to be used in diagnostics and further research.

A-173 Evaluation of Artificial Intelligence (AI)-assisted Flow Cytometry Analysis for Plasma Cell Neoplasms

Abstract Background Flow cytometry plays a critical role in diagnosing hematologic disorders, yet manual analysis is time-consuming and prone to variability. Significant advancements in artificial intelligence (AI) within healthcare have occurred in recent years. Previously, we developed and evaluated an AI-assisted automated system for diagnosing acute leukemia. This study aims to assess the effectiveness of the AI system in diagnosing plasma cell neoplasms. Methods A total of 76 bone marrow samples were analyzed using a panel of antibodies (CD38, CD56, CD138, Kappa, and Lambda) to determine plasma cell percentages and immunophenotype. All samples were used for initial diagnosis of plasma cell neoplasm or therapeutic monitoring. Manual analysis using Kaluza™ software served as the gold standard. The same flow cytometry data was processed by the AI software (DeepFlow™) for comparison. Pearson correlation coefficients were used to compare AI and manual analysis for diagnosis and plasma cell count. Results Among the 76 cases, manual analysis identified 57 positive cases for monoclonal plasma cells (36 cases of CD38+/CD138+/Kappa+ and 21 cases of CD38+/CD138+/Lambda+), with 89% showing aberrant CD56 expression. AI identified 49 monoclonal cases, missing 8 cases with low plasma cell levels (&amp;lt;0.07%) or brighter fluorescence. AI accurately matched 97% of immunophenotypes with manual analysis. Pearson correlation coefficients demonstrated excellent correlation (r=1.0) between manual and AI plasma cell percentages. Furthermore, AI has also enhanced laboratory efficiency, reducing analysis time per case from 10 minutes with manual methods to just 3 minutes with AI analysis. Conclusions This study shows promise for AI-assisted flow software in diagnosing plasma cell neoplasms. AI has enhanced operational efficiency by reducing the manual processing and analysis time. Further training is needed to improve the identification of variations in fluorescence intensity and low plasma cell levels.

Altered B-cell, plasma cell and antibody immune profiles in blood of systemic mastocytosis

Systemic mastocytosis (SM) is a heterogeneous disease characterised by an expansion of KIT-mutated constitutively activated mast cells (MC) which release MC mediators that might act on the tumour microenvironment including other immune cells. Here we investigated the blood distribution of B-cell, plasma cell (PC) and antibody-isotype compartments in SM. We used spectral flow cytometry and the EuroFlow Immunomonitoring panel and Lymphocyte Screening Tube to quantify B-cells, PC and their subsets in blood of 108 SM patients - 35 bone marrow mastocytosis (BMM), 64 indolent SM (ISM), 9 aggressive SM (ASM)- vs 117 age-matched healthy donors (HD) and paired bone marrow (BM) samples of 31 SM vs 17 controls, respectively. In parallel, immunoglobulin (Ig) M, IgD, IgG, IgA and IgE plasma levels of were measured. Compared to HD, SM patients showed an increased immature B-cell production in BM (P=0.003) associated with a greater release of pre-germinal center immature (P<0.001) and naive CD5+ B-lymphocytes (P<0.001) to blood, but a pronounced decrease in PC counts of all different IgH-isotypes and subclasses (P≤0.001) together with overall increased IgM (P=0.001) and IgD (P<0.001) plasma levels. Of note, different immune profiles were found per diagnostic subtype of the disease with progressively greater counts in blood of immature B-lymphocytes together with decreased IgMD+, IgG2+, IgA1+ and IgA2+ MBC (P≤0.032) and elevated IgM (P=0.017) plasma levels in ASM cases, increased IgM (P=0.001) and IgD (P=0.001) plasma levels in ISM patients and exacerbated IgE (P<0.001) with decreased IgG (P=0.008) plasma levels in BMM cases. Our results reveal a significant dysregulation of the B-cell and PC compartments in blood of SM patients, consistent with distinctly altered antibody-isotype profiles in plasma of BMM vs ISM vs ASM patients.

Beyond diagnosis: exploring the significance of IgG4+ plasma cell count through immunostaining in IgG4-related disease.

To evaluate whether the grade of IgG4+ plasma cell infiltration in biopsies is associated with clinical or serologic outcomes in IgG4-RD. We included 57 patients with biopsy proven IgG4-RD according to the Comprehensive Diagnostic Criteria and/or the 2019 ACR/EULAR Classification Criteria. We collected histological, clinical (disease duration, phenotype, remission and relapses) and serological variables. 29 (50.9%) patients were men, mean age 49.9 years, with a median disease duration of 22 months. The distribution among clinical phenotypes were 14% pancreato-hepato-biliary, 12.3% retroperitoneal/aortic, 29.8% head and neck-limited, 29.8% Mikulicz/systemic and 14% undefined. Thirty-nine patients had a proliferative and 18 a fibrotic phenotype. Most biopsies were from lacrimal gland, lymph node, pancreas, orbit, kidney, retroperitoneum and thyroid gland. Thirty-nine (68.4%) patients had <100 IgG4+ plasma cells/HPF and 18 (31.6%) ≥100 IgG4+ plasma cells/HPF. Patients with ≥100 IgG4+ plasma cells/HPF were more likely to belong to the pancreato-hepato-biliary and the proliferative phenotypes, had fewer relapses and a higher remission rate. On multivariate analysis, the OR for remission at last follow-up was 6.7, 95% CI 1.1-4.42, p=0.03. The log-rank test showed a difference in relapse-free survival between the two groups (HR 2.6, 95% CI 1.2-5.6, p=0.01). According to the ROC analysis, patients with more than 61 IgG4+ plasma cells were less likely to relapse. A count of ≥100 IgG4+ plasma cells/HPF may identify patients with a proliferative phenotype, fewer relapses and a higher remission rate.

Population pharmacokinetic study and application of ticagrelor and AR-C124910XX after percutaneous coronary intervention in Chinese patients with acute coronary syndrome.

This study aimed to understand the pharmacokinetics of ticagrelor in Chinese patients with acute coronary syndrome (ACS), identify influencing factors, and inform ticagrelor treatment optimization. Data from 195 ACS patients, including 491 plasma ticagrelor concentration timepoints and clinical information, were analyzed using NONMEN for pharmacokinetic (PK) parameter factors. The model underwent internal validation with bootstrap methodology. The PK curve of ticagrelor was well delineated using a one disposition compartment model with first-order absorption rate constant, 0.67/h. When the direct bilirubin levels and white plasma cell counts increased, female patients showed decreased glomerular filtration rate, decreased ticagrelor clearance rate, and increased exposure. When the direct bilirubin levels increased and body weight and hemoglobin decreased, rs6787801 was GG compared with AA and GA, the ticagrelor metabolite clearance rate decreased and exposure increased. The study offers key insights into ticagrelor's dose-exposure relationship post-percutaneous coronary intervention in ACS patients, highlighting factors critical for personalized treatment strategies.

Increased levels of regulatory T cells and IL-10-producing regulatory B cells are linked to improved clinical outcome in Parkinson's disease: a 1-year observational study.

Whilst the contribution of peripheral and central inflammation to neurodegeneration in Parkinson's disease and the role of the immune response in this disorder are well known, the effects of the anti-inflammatory response on the disease have not been described in depth. This study is aimed to assess the changes in the regulatory/inflammatory immune response in recently diagnosed, untreated PD patients and a year after. Twenty-one PD patients and 19 healthy controls were included and followed-up for 1 year. The levels of immunoregulatory cells (CD4+ Tregs, Bregs, and CD8+ Tregs); classical, nonclassical, and intermediate monocytes, and proinflammatory cells (Th1, Th2, and Th17) were measured by flow cytometry. Cytokine levels were determined by ELISA. Clinical follow-up was based on the Hoehn & Yahr and UDPRS scales. Our results indicate that the regulatory response in PD patients on follow-up was characterized by increased levels of active Tregs, functional Tregs, TR1, IL-10-producing functional Bregs, and IL-10-producing classical monocytes, along with decreased counts of Bregs and plasma cells. With respect to the proinflammatory immune response, peripheral levels of Th1 IFN-γ+ cells were decreased in treated PD patients, whilst the levels of CD4+ TBET+ cells, HLA-DR+ intermediate monocytes, IL-6, and IL-4 were increased after a 1-year follow-up. Our main finding was an increased regulatory T cell response after a 1-year follow-up and its link with clinical improvement in PD patients. In conclusion, aftera 1-year follow-up, PD patients exhibited increased levels of regulatory populations, which correlated with clinical improvement. However, a persistent inflammatory environment and active immune response were observed.

Comparison of plasma cell bone marrow counts by different methods in patients diagnosed with plasma cell disorders

IntroductionPlasma cell quantification in bone marrow is important for diagnosis, prognosis, and treatment of plasma cell diseases. It can be performed by several methods such as aspiration, imprint and flow cytometry, and biopsy. ObjectivesTo compare plasma cell counts at diagnosis of plasma cell diseases using different methods. MethodsAn observational study was carried out of laboratory results of adult patients with plasma cell diseases, who underwent aspiration, imprint cytology, flow cytometry (CD38, C138) and biopsy in a single institution between January 2015 and May 2021. The intraclass correlation coefficient was used to assess agreement between different methods with results stratified into three groups: <10%; 10–59% and ≥60% of infiltration. ResultsSixty-seven cases were studied: 59.7% were men with a median age of 70 (range: 32–85) years. The diagnoses were multiple myeloma in 61%, gammopathy of undetermined significance in 25.4%, smoldering myeloma in 6% and other plasma cell dyscrasias in 7.6%. Less than 10% infiltration was found in 32 (47.7%), 35 (52.2%), 44 (65.7%) and 25 (37.3%) of patients, respectively by aspiration, imprint cytology, flow cytometry and biopsy. Infiltration ≥60% was detected in 7 (10.4%), 4 (6.0%), 2 (3.0%) and 21 (31.3%) cases, respectively. There was disagreement between the results in 37 (55.2%) of patients. Of these, 28 had greater infiltration in biopsies. The concordance (Kappa index) of biopsy with aspiration, imprint and flow cytometry was 0.501, 0.408 and 0.17; of aspiration with imprint and flow cytometry, it was 0.738 and 0.541 and between imprint and flow cytometry, it was 0.573%. ConclusionsOnly aspiration and imprint cytology results agreed. Biopsy showed greater infiltrations than the other methods, but aspiration, and imprint and flow cytometry provided additional data in the diagnosis and thus should also be performed.

Leucémie à plasmocytes secondaire : à propos d’un cas rare et revue de littérature Secondary plasma cell leukemia: a rare case report and review of the literature

Plasma cell leukemia is a hematological malignancy characterized by monoclonal proliferation of plasma cells in the peripheral blood and bone marrow. It is defined by a blood plasma count greater than 2 × 109/L (2G/L) and/or the presence of more than 20% plasma cells in the white blood cell count. There are two forms: the primary or de novo form, which manifests itself immediately as leukemia, and the secondary form , which corresponds to the evolution of a known multiple myeloma. It is a highly aggressive form of plasma cell neoplasm, with a poor prognosis despite therapeutic advances. In this context, we present a rare case of secondary plasma cell leukemia in a 56-year-old female patient, diagnosed at the hematology laboratory of the Hassan II University Hospital in Fez. The patient was being followed for multiple myeloma, for which she was put on chemotherapy according to the Velcade + Thalidomide + Dexamethasone protocol. Six months later, the patient presented with a febrile hemorrhagic syndrome. The blood count showed hyperleukocytosis at 39.4 G/L and bicytopenia (Hemoglobin = 7.9 g/dl, Platelets = 58 G/L). Blood smear showed a 36% plasma cell count suggestive of secondary plasma cell leukemia. At the end of this observation, we underline the crucial importance of regular monitoring by careful cytological examination of the blood smear in all patients followed for multiple myeloma.

P-302 Oral ofloxacin combined with vaginal metronidazole for chronic endometritis: a prospective open-label cohort study

Abstract Study question Does oral ofloxacin 2x200 mg for 10 days combined with vaginal metronidazole 1x500 mg for 10 days reduce the activity of chronic endometritis? Summary answer The investigated antibiotics may be beneficial in some cases of chronic endometritis, except in cases of concomitant endometrial polyp, where they adversely affect treatment outcome. What is known already Chronic endometritis (CE) is characterized by the presence of atypical plasma cell infiltrates (CD138 positive) in the endometrial stroma. CE may adversely affects fertility by reducing endometrial receptivity, impairing decidualization and uterine contractility, thus increasing the risk of recurrent pregnancy loss and implantation failure. Some idiopathic infertility cases are attributed to CE. The diagnostics of CE is challenging because the clinical evaluation is nonspecific and the immunohistochemical criteria are not consistent between research centers. Literature data suggested that successful treatment of CE with antibiotics could improve live birth rates, but a preferred treatment regimen has not been identified. Study design, size, duration A prospective cohort study included women undergoing office hysteroscopy (OH) at a tertiary referral center in 2021-2023. Women diagnosed with CE were assigned, based on their preference, to the intervention arm of ofloxacin 2x200 mg orally for 10 days + metronidazole 500 mg vaginally for 10 days or to the control arm (no antibiotics). 100 women underwent repeated hysteroscopy after 3 cycles, of which 77 received antibiotics and 23 were controls. Participants/materials, setting, methods Women aged 18-45 who met eligibility criteria for OH were included. Karl Storz 5.0 mm Bettocchi® operating sheath with a 2.9 mm 30° telescope and a 5Fr working channel was used. Endometrial lesions or biopsy specimens were collected with grasping forceps, blunt scissors or bipolar needle electrode. Saline was used as the dilating medium. Tissue samples were subjected to histopathological examination. Plasma cells were detected by immunohistochemical staining with Monoclonal Mouse Anti-Human CD138 antibodies. Main results and the role of chance The most common findings identified at hysteroscopy were endometrial polyps (43%), polypoid endometrium (38%), micropolyps (23%), focal (27%) and diffuse hyperemia (18%) and cesarean scar defect (14%). CE was defined as the presence of &amp;gt; 2 plasma cells/mm2. The Mann-Whitney U test showed a significantly higher plasma cell count (PCC) in the presence of a cesarean scar defect than in other cases (23 vs. 18/mm2, p = 0.031) and a tendency towards a lower PCC in micropolyps (9 vs. 21/mm2, p = 0.063). The correlation of hysteroscopic findings with CE activity was weaker than expected. Using repeated measures ANOVA, no difference in PCC between primary and follow-up hysteroscopy was found in either arm (p = 0.8), while post-hoc tests showed that the mean PCC decreased from 12 to 5/mm2 (p = 0.02) in the intervention arm, and from 14 to 6 (p = 0.23) in the control arm. On follow-up hysteroscopy, polyp remission was observed significantly more often in the control arm than in the intervention arm (61% vs. 27%, pChi2=0.02), in which de novo polyp was observed in 4%. In polypoid endometrium, the remission rate was similar in both arms (19% vs. 22%), and the rate of de novo lesions was significantly lower in interventional arm (16% vs. 39%, pChi2=0.035). Limitations, reasons for caution The limitations include the small sample size, the CE definition based on the number of plasma cells calculated using the Receiver Operating Characteristic curve for the purposes of the study, and the empirical selection of antibiotic therapy. Wider implications of the findings Due to the weak correlation of hysteroscopic findings with CE activity, the diagnostics and treatment of CE based on clinical evaluation of the uterine cavity is not justified. Poorly selected antibiotic therapy may adversely affect the outcomes of treatment. Trial registration number 1072.6120.322.2020

Reference intervals for bone marrow cells in juvenile and young adult cats bone marrow cytology.

The current bone marrow (BM) reference intervals (RI) are based on a limited number of cats. Age-related changes in BM variables might be important,possibly affecting the interpretation of the results. Establish BM cytologic reference intervals (RIs) and association of age and sex on these findings, in healthy juvenile and young adult cats. BM aspirates of cats deemed healthy based on history and clinical, CBC, serum chemistry findings, and negative retrovirus serology were obtained and examined cytologically. The examination included a 1000-nucleated differential cell count and cell morphologic assessment. RIs were calculated using parametric, robust, and nonparametric methods. The cytologic findings were examined for associations with sex and age. The study included 40 cats (females, 22 [55%]; males, 18 [45%]) with a median age of 1.5 years (range 0.5-5). Most calculated RIs were similar to those previously reported. BM plasma cell and monocyte counts were weakly and positively correlated with age (rs, .312 and .373, respectively; P < .05). Metarubricytes were higher infemales (mean, 25.1%; SD, 6.0%) than males (mean, 21.2%; SD, 6.0%; P < .05). The BM differential cell counts determined in this study can serve as RIs for cats aged 0.5-5 years.

Plasma proteome signature of canine acute haemorrhagic diarrhoea syndrome (AHDS).

Acute haemorrhagic diarrhoea is a common complaint in dogs. In addition to causes like intestinal parasites, dietary indiscretion, intestinal foreign bodies, canine parvovirus infection, or hypoadrenocorticism, acute haemorrhagic diarrhoea syndrome (AHDS) is an important and sometimes life-threatening differential diagnosis. There is some evidence supporting the link between Clostridium perfringens toxins and AHDS. These toxins may be partially responsible for the epithelial cell injury, but the pathogenesis of AHDS is still not fully understood. Recent studies have suggested that severe damage to the intestinal mucosa and associated barrier dysfunction can trigger chronic gastrointestinal illnesses. Besides bloodwork and classical markers for AHDS such as protein loss and intestinal bacterial dysbiosis, we focused mainly on the plasma-proteome to identify systemic pathological alterations during this disease and searched for potential biomarkers to improve the diagnosis. To accomplish the goals, we used liquid chromatography-mass spectrometry. We compared the proteomic profiles of 20 dogs with AHDS to 20 age-, breed-, and sex-matched control dogs. All dogs were examined, and several blood work parameters were determined and compared, including plasma biochemistry and cell counts. We identified and quantified (relative quantification) 207 plasmatic proteins, from which dozens showed significantly altered levels in AHDS. Serpina3, Lipopolysaccharide-binding protein, several Ig-like domain-containing proteins, Glyceraldehyde-3-phosphate dehydrogenase and Serum amyloid A were more abundant in plasma from AHDS affected dogs. In contrast, other proteins such as Paraoxonase, Selenoprotein, Amine oxidases, and Apolipoprotein C-IV were significantly less abundant. Many of the identified and quantified proteins are known to be associated with inflammation. Other proteins like Serpina3 and RPLP1 have a relevant role in oncogenesis. Some proteins and their roles have not yet been described in dogs with diarrhoea. Our study opens new avenues that could contribute to the understanding of the aetiology and pathophysiology of AHDS.

Improved Outcome of Primary Plasma Cell Leukemia in the Current Era with the Use of Novel Agents and Autologous Bone Marrow Transplants-A Single Centre Experience.

Primary Plasma cell leukemia (pPCL) is an aggressive variant of plasma cell dyscrasias. Diagnostic criteria of plasma cell leukemia were recently updated by international myeloma working group to with more than 5% circulating plasma cells or absolute plasma cell count of more than 500/µL. We performed a retrospective analysis of patients diagnosed with pPCL in our department from 2017 to 2022. Clinical characteristics including the symptoms at presentation, organomegaly, bony involvement and extramedullary involvement were collected. Laboratory parameters including the biochemistry serum protein electrophoresis, serum immunofixation, serum free light chain assay, immunoglobulin profile were sent. Treatment and follow up data was collected. Fifteen patients were diagnosed (8 females and 7 males), median age 59 years (34-70). Six were lost to follow up and nine patients who received treatment at our hospital were analyzed for survival outcome. First line treatment was bortezomib- dexamethasone and immunomodulatory drugs (IMiD). Six (66%) achieved partial response or more and 3 had progressive disease. Five of the nine patients (55%) underwent autologous transplantation. Two out of 5 patients (40%) in the transplant group and 3 of the 4 patients (75%) in the non transplant group have died of the progressive disease. Overall survival was 45% at a median follow up of 14 months. Median OS for patients who underwent auto SCT was 16 months (12-22) versus 10 months (8-12) for patients who did not undergo transplant (Student t test; p value 0.018). Three of the patients achieved MRD negativity after transplant and post transplant consolidation therapy. Survival appears to be improved in patients who respond to initial therapy and are able to achieve MRD negativity which should be the goal of treatment in these patients.

Whole-Genome Sequencing for Copy Number Abnormalities in Multiple Myeloma Supersedes Karyotype Analysis and Fluorescent in Situ Hybridization

Whole-Genome Sequencing for Copy Number Abnormalities in Multiple Myeloma Supersedes Karyotype Analysis and Fluorescent in Situ Hybridization Jiali Li 1, Shaobing Gao 2, Zhenling Li 3, Yinyin Chang 4, Xiangxiang He 4, Yuexin Cheng 5, Manqian Li 4, Chunxiao Yao 4, Shiyong Li 6, Dandan Zhu 4, Shichun Tu 4, Mao Mao 6,7, #, Xi Zhang 1, #, Li Gao 1, # 1 The Xinqiao Hospital of Third Military Medical University, Chongqing 214426, China 2The Affiliated Cancer Hospital of Zhengzhou University and Henan Cancer Hospital, Zhengzhou 450003, China 3China-Japan Friendship Hospital, Beijin100029,China 4Clinical Laboratories, Shenyou Bio, Zhengzhou 450000, China 5Yancheng No. 1 People's Hospital, The Yancheng Clinical College of Xuzhou Medical University, Yancheng 224006, China 6Research &amp; Development, SeekIn Inc., Shenzhen 518000, China 7Yonsei Song-Dang Institute for Cancer Research, Yonsei University, Seoul 03722, Korea Corresponding authors. Tel.: +86-15601973722(Maomao); 13808310064(Xi Zhang) 13228686076(Li Gao) Corresponding authors Email: mao_m@yahoo.com (Maomao); zhangxxi@sina.com (Xi Zhang); gaotiantiantiger@163.com (Li Gao) Introduction Prognosis and management of multiple myeloma (MM) relies on risk stratification based on copy number abnormalities (CNA) detection results. CNA is conventionally evaluated by cytogenetic methods including karyotyping and flurescence in situ hybridization (FISH), both limited by the invasiveness, low proliferative activity of plasma cell and the low plasma cell count. LeukoPrint is a novel shallow whole-genome sequencing (sWGS) based approach to profile CNA in bone marrow cells and/or circulating cell-free DNA (cfDNA). We compared the CNA detection results by LeukoPrint, karyotyping and FISH using bone marrow or plasma samples from multiple myeloma patients. Methods A total of 128 patients were enrolled in this study. Karyotyping and FISH were performed by conventional approaches. CNA was detected by sWGS in genomic DNA from bone marrow aspiration samples and CD138+ enriched plasma cells as well as in plasma cfDNA. CNA degree was measured by log R ratio. Significant mutational peak region was identified by GISTIC2. Risk stratification was performed following the Mayo Stratification of Myeloma and Risk-adapted Therapy (mSMART) criteria. Results In this study, 908 CNAs were identified by sWGS in 96 (75.0%, 96/128) patients, while the positive rate of CNA detected by karyotyping and FISH was 7.9% and 56.7% , respectively. A total of 368 whole chromosome aberration events were identified by sWGS, 66.0% (243/ 368) of which were trisomy that defined 39.6% (38/96) of patients as hyperdiploidy and the remaining 60.4% (58/96) as non-hyperdiploidy. The most frequently mutational CNA peak region were amp(1q21), del(1p), del(6q), del(8p), del(13q14) and del(14q). The concordance rate between sWGS and FISH in detecting the four conventional loci (1q21 gain, 13q14 del, 13q14.3 del, and 17p13 del) was 81.8%. Comparing to the conventional approaches, sWGS provided new CNA information for 75.0% of the patients, and redefined the risk stratification for 17.2% of patients according to mSMART criteria. CNA detection results using paired bone marrow aspiration samples and enriched plasma cell samples from 8 patients were compared. We found that more CNAs (81 vs 58) and higher Log R ratio of CNAs were detected using enriched plasma cells than that using bone marrow aspiration samples. The concordance rate of the CNA detection results by sWGS between paired bone marrow genomic DNA and plasma cfDNA from another 9 patients was 83.3%. Conclusions LeukoPrint is an automated, convenient and cost-effective approach to depict CNA profile in genomic DNA or cfDNA. This method is superior to conventional approaches when used for CNA testing, and the practice of this method could improve prognostic stratification of MM patients.

Increased IgG4+ plasma cells are common in excised lymph nodes from children and adolescents without IgG4-related disease.

Lymphadenopathy is a common finding in patients with IgG4-related disease (IgG4-RD) and often associated with increased IgG4+ plasma cells in this setting. The histologic features of so-called IgG4-related lymphadenopathy (IgG4-LAD) have seldom been investigated in children and adolescents, and step-wise progression to extranodal IgG4-RD has not been described. This study was performed to further evaluate the frequency, pathologic features, and clinical significance of IgG4-LAD-like histologic changes in the pediatric setting. We analyzed 37 benign lymph nodes collected semi-consecutively from children aged 0-18 years at our institution for both absolute and relative IgG4+ plasma cell abundance and recurrent histomorphologic patterns associated with IgG4-LAD. The combination of IgG4+/IgG+ plasma cell ratio >40% and IgG4+ plasma cell count ≥50 were considered as IgG4-LAD-like per expert consensus guidelines. Seven cases (19%) met both diagnostic criteria. The dominant histomorphologic patterns were follicular hyperplasia (n = 6), interfollicular expansion (n = 3), and progressive transformation of germinal centers (n = 3). Extranodal manifestations of IgG4-RD were not identified in this cohort (38 months average follow-up). Instead, clinical and laboratory findings indicated that lymph node enlargement in most patients could likely be attributed to alternative processes including antecedent dentistry, concurrent infection, and incipient Crohn's disease. Our findings suggest that the histologic features of IgG4-LAD are likely much more common in children and adolescents than previously recognized, often existing in complex with common reactive lymphadenopathies. The diagnostic value of routine immunohistochemical assessment for IgG4+ plasma cells in benign lymph nodes from pediatric patients without established extranodal IgG4-RD and/or other supportive clinical and laboratory data is therefore uncertain.

Immune cell kinetics and antibody response in COVID-19 patients with low-count monoclonal B-cell lymphocytosis.

Low-count monoclonal B-cell lymphocytosis (MBLlo ) has been associated with an underlying immunodeficiency and has recently emerged as a new risk factor for severe COVID-19. Here, we investigated the kinetics of immune cell and antibody responses in blood during COVID-19 of MBLlo versus non-MBL patients. For this study, we analyzed the kinetics of immune cells in blood of 336 COVID-19 patients (74 MBLlo and 262 non-MBL), who had not been vaccinated against SARS-CoV-2, over a period of 43 weeks since the onset of infection, using high-sensitivity flow cytometry. Plasma levels of anti-SARS-CoV-2 antibodies were measured in parallel by ELISA. Overall, early after the onset of symptoms, MBLlo COVID-19 patients showed increased neutrophil, monocyte, and particularly, plasma cell (PC) counts, whereas eosinophil, dendritic cell, basophil, and lymphocyte counts were markedly decreased in blood of a variable percentage of samples, and with a tendency toward normal levels from week +5 of infection onward. Compared with non-MBL patients, MBLlo COVID-19 patients presented higher neutrophil counts, together with decreased pre-GC B-cell, dendritic cell, and innate-like T-cell counts. Higher PC levels, together with a delayed PC peak and greater plasma levels of anti-SARS-CoV-2-specific antibodies (at week +2 to week +4) were also observed in MBLlo patients. In summary, MBLlo COVID-19 patients share immune profiles previously described for patients with severe SARS-CoV-2 infection, associated with a delayed but more pronounced PC and antibody humoral response once compared with non-MBL patients.

Factors affecting the features of platelet-rich plasma in patients with knee osteoarthritis.

The aim of this study was to present an analysis of platelet-rich plasma obtained from patients with knee osteoarthritis and reveal the factors affecting its features. A total of 62 patients (mean age: 56.68 ± 7.13 years) with symptomatic knee osteoarthritis were included in this study. Age (years), gender, height (m), weight (kg), body mass index (kg/m2), duration of symptoms, smoking status, smoking index, general health status, and physical activity scores were recorded. Whole blood and platelet-rich plasma cell counts were performed with a hematology analyzer. White blood cell, red blood cell, and platelet counts were recorded. According to the dose of injected platelets, efficiency of the procedure, purity of platelet-rich plasma, and activation classification, dose of platelets, efficiency of the procedure (platelet recovery rate, %), and purity of the obtained platelet-rich plasma product (relative composition in platelets, %) were calculated. Correlation analysis between the features of platelet-rich plasma and the patient-related variables, including age, gender, body mass index, smoking status, smoking index, presence of other health conditions, physical activity scores, duration of symptoms, and pain levels, was performed. Dose of injected platelets, efficiency of the procedure, purity of platelet-rich plasma, and activation analysis showed that the dose of injected platelets was 3.25 billion, the efficiency of the process was 77%, and the purity rate of the platelet-rich plasma was 98.4%. Platelet-rich plasma platelet count was correlated with whole blood platelet count (r = 0.81, P < .001), whole blood white blood cell count (r = 0.39, P = .002), smoking status (r = 0.56, P = .03), smoking index (r = -0.63, P = .002), and the presence of hypertension (r = -0.31, P=.04). Platelet-rich plasma white blood cell and purity of platelet-rich plasma were correlated with the smoking status of the patients (r = 0.52, P = .01; r = 0.64, P = .003, respectively). This study has demonstrated that high dose and very pure platelet-rich plasma with medium efficiency was yielded with this platelet-rich plasma preparation procedure; whole blood platelet count, the presence of hypertension, and the smoking status of patients affect the features of the obtained platelet-rich plasma. Level IV, Diagnostic Study.

The role of IgG4-positive plasma cell population in classic Hodgkin lymphoma.

The effect of IgG4, which constitutes the least of the IgG subclasses, on the pathogenesis and prognosis of lymphoma or solid tumors is one of the research topics of interest in recent years. The role of IgG4, which has been reported to suppress antitumor immunity, in classic Hodgkin's lymphoma (cHL), which is recognized by its pathognomonic microenvironment, is not yet clearly known.The aim of this study was to determine IgG4-positive plasma cell density in the cHL microenvironment and to compare it with histopathological and clinical parameters. In addition, the role of the increase in IgG4-positive cells in the development of relapse after treatment was also investigated.A retrospective cross-sectional study.Ninety-four patients with the initial diagnosis of cHL who had no comorbidity or no treatment history and forty-one reactive lymph nodes with follicular hyperplasia findings were included in the study. Three hot-spot areas were identified with reference to the IgG4 sections. Mean IgG4-positive plasmacyte counts and IgG4/IgG ratios were determined and compared with histopathological characteristics.The mean IgG4 + plasma cell count was 33.57 in cHL cases and 47.04 in the control group (p = 0.233). IgG4/IgG ratio was significantly higher in cHL compared with the control group (0.27 vs. 0.21, p = 0.021). The IgG4/IgG ratio was found to be higher in younger patients with classic Hodgkin lymphoma, with a low correlation (p = 0.028, r = - 0.226). There was no relationship with gender, lymph node location, histological subtype, EBV positivity and bone marrow infiltration. It was observed that IgG4/IgG ratio was higher in early-stage patients (p = 0.022). No significant IgG4 + cell increase was detected in the initial diagnosis and relapse slides of six patients who developed relapse after standard treatment, resulting in a cure.Novel therapeutic modalities targeting microenvironmental components have been reported to show dramatic effects, particularly in relapsed or refractory patients. Detailed characterization of the cHL inflammatory milieu will be useful for the identification of alternative targets. IgG4 subclass antibodies, which have been described to have anti-inflammatory effects, may have prognostic significance in a proportion of cHL patients.

IgG4-Related Disease, a Systemic or Local Disease: This is the Problem

Background: Immunoglobulin G4-related disease (IgG4-RD) is a fibroinflammatory condition that forms tumefactive lesions in various organs, presenting with diverse clinical manifestations. Diagnosis of IgG4-RD could be challenging and relies on characteristic histopathological findings and elevated IgG4+ plasma cell counts. Case Presentation: The patient was a 48-year-old woman with diabetes mellitus initially presented with a painless breast mass. Imaging indicated irregular opacities and lymph nodes in the breast. Biopsy results revealed lymphoplasmacytic infiltration with IgG and IgG4 positivity, overall compatible with IG4-related disease. A mass resection was performed, and intraoperative assessment ruled out malignancy. Then, with a 5-month interval of no symptoms, the patient developed a cheek mass, initially misdiagnosed as low-grade spindle cell sarcoma. Subsequently, she underwent partial maxillectomy and further examination and histology confirmed IgG4-RD, meeting all diagnostic criteria. Conclusion: IgG4-RD is a rare, systemic fibroinflammatory condition with potential to mimic neoplastic processes. While IgG4 serum levels can be elevated, they are not a reliable diagnostic criterion. The two main features of IgG-related disease are an elevated number of IgG4-positive plasma cells within the tissue and a characteristic histopathological appearance including a dense lymphoplasmacytic infiltrate, storiform fibrosis, and obliterative phlebitis. Overall, diagnosis is based on the balance of clinical features, histopathology, and serum markers. This case underscores the diagnostic challenges posed by IgG4-RD, often mistaken for malignancies due to its invasive behavior and emphasizes the importance of considering IgG4-RD in differential diagnoses for tumor-like lesions, particularly when affecting multiple organs, to enable timely treatment decisions and prevent unnecessary interventions.

Metabolic Syndrome, Hyperlipidemias, and Associated Clinical Markers Among Military Airmen

BACKGROUND: Due to the increased overweight and obesity prevalence in Colombia, the aim of this study is to determine the frequency of metabolic syndrome (MetS) among Colombian Air Force military aviators, crews, remote piloted aircraft operators, and air traffic controllers and relationships with clinical markers. METHODS: Operationally active Columbian Air Force military personnel examined periodically at the Colombian Air Force Aerospace Medical Directorate were chosen for a cross-sectional study performed among 2179 subjects. Medical history, anthropometrics, and laboratory results were analyzed and frequencies, correlations, and odds ratios were calculated. RESULTS: Overall prevalence of MetS was 21.7%; in subjects with BMI ≥25, frequency increased to 36% vs. those with BMI &lt;25. Hypertriglyceridemia was present in 31%, impaired fasting glucose 12.5%, hypertension (≥130/85 mmHg) 14.4%, low HDL-C 35.2%, and increased waist circumference 46.2%. Those with three criteria were 14.6%, four criteria 5.8%, and five criteria 1.2%. Pilots had a significantly lower prevalence of MetS at 17.7% and an adjusted OR of 0.61 (0.49–0.76) than other crew; hyperuricemia was three times more likely (3.2–5.1) and hypercholesterolemia OR was 2.3 (1.9–2.9). Subjects with MetS had a significantly higher fat percentage, waist circumference, low-density lipoprotein, very low-density lipoprotein, non-high-density lipoprotein cholesterol, atherogenic index of plasma (AIP), uric acid, and white blood cell (leukocyte) count, and a lower estimated glomerular filtration rate. There is a linear relationship of the AIP and waist circumference, BMI, uric acid, and white blood cell count. DISCUSSION: MetS prevalence among Colombian Air Force aviators is lower than the general population, higher than other countries, and displays worse lipid profiles that increase cardiovascular and diabetes mellitus risk within the military. Malpica D. Metabolic syndrome, hyperlipidemias, and associated clinical markers among military airmen. Aerosp Med Hum Perform. 2023; 94(8):604–609.