P090 Rheumatic symptoms as the first manifestations of underlying systemic disorders

Rheumatology ◽  
2021 ◽  
Vol 60 (Supplement_5) ◽  
Author(s):  
Megha Unadkat ◽  
Angela Migowa

Abstract Background Joint and limb pain is a common presenting complaint within the paediatric population, affecting 10–20% of school-age children. They are also a major reason for referral to paediatric rheumatology clinics (1). The differential diagnosis for joint pain and/or swelling can include both benign and malignant causes. Studies have found that < =1% of musculoskeletal complaints are caused by neoplasia, mostly acute lymphoblastic leukaemia (ALL) (1). It is essential to exclude life-threatening conditions such as infections and malignancies before initiation of treatment for presumptive Juvenile Idiopathic Arthritis (JIA) (2). The objective of this case series is to highlight the musculoskeletal features and the clinical course among children with haematological malignancies. Methods Charts of patients referred from rheumatology clinic to the haemato-oncology clinic were selected. A detailed chart review was then carried out to obtain information on clinical presentation, diagnostic work up and clinical progress of the patient. Results Case 1 CS, A 2 year and 2 months old girl who presented to paediatric rheumatology clinic with 5 month history of lower limb pain associated with a fever of 40’C. The pain progressively worsened leading to inability to walk. She had previously been treated for Idiopathic Thrombocytopenic Purpura with steroids. Initial Bone Marrow Aspiration (BMA) had ruled out leukaemia. Four months later, her condition had worsened and was referred to our facility where a peripheral blood film and flow cytometry confirmed B cell ALL. Treatment was initiated but unfortunately the patient’s condition deteriorated and eventually passed away. Case 2 NM, a 2 year and 10 months old girl presented to our paediatric rheumatology clinic with bilateral foot pain for 8 weeks. She was initially limping and progressed to inability to walk within a month. Rheumatology review in Tanzania diagnosed the patient as JIA and started on steroids. On examination, systemic findings were normal except bilateral ankle joint effusion and tenderness. An X-ray displayed bilateral transverse lucencies in the metaphyseal ends of the tibia and fibula. BMA was done and confirmed ALL. Treatment was initiated and the patient attained remission after 18 months of therapy. She remains well 2 years after completion of therapy. Case 3 NC, a 4-year-old girl, presented with multiple joint pains for 2 weeks. Pain was associated with night awakening and recurrent fevers. C-Reactive Protein (CRP) was persistently high and was treated with several courses of antibiotics. She was assessed by a haemato-oncologist with unremarkable findings. A decision was made to do a BMA and flow cytometry, which confirmed B cell ALL. Conclusion Musculoskeletal complaints are frequently the initial manifestations of acute leukaemia in childhood (60%) and should be considered in the differential diagnosis of JIA (3). Initially laboratory test results can be normal therefore it is necessary to follow these children closely and request serial tests, until there is a clearly defined diagnosis of neoplasia. Finally, malignancy should be eliminated before introduction of immunosuppressive therapy as the use of steroids may mask and delay its diagnosis (2).

Author(s):  
Sebastian Böttcher ◽  
Robby Engelmann ◽  
Georgiana Grigore ◽  
Paula Carolina Fernandez ◽  
Joana Caetano ◽  
...  

Reproducible expert-independent flow-cytometric criteria for the differential diagnoses between mature B-cell neoplasms are lacking. We developed an algorithm-driven classification for these lymphomas by flow cytometry and compared it to the WHO gold standard diagnosis. Overall, 662 samples from 662 patients representing nine disease categories were analyzed at 9 laboratories using the previously published EuroFlow 5-tube-8-color B-cell chronic lymphoproliferative disease antibody panel. Expression levels of all 26 markers from the panel were plotted by B-cell entity to construct a univariate, fully standardized diagnostic reference library. For multivariate data analysis we subsequently utilized Canonical Correlation Analysis of 176 training cases to project the multi-dimensional space of all 26 immunophenotypic parameters into 36 two-dimensional plots for each possible pair-wise differential diagnosis. Diagnostic boundaries were fitted according to the distribution of the immunophenotypes of a given differential diagnosis. A diagnostic algorithm based on these projections was developed and subsequently validated using 486 independent cases. Negative predictive values exceeding 92.1% were observed for all disease categories except for follicular lymphoma. Particularly high positive predictive values were returned in chronic lymphocytic leukemia (99.1%), hairy cell leukemia (97.2%), follicular lymphoma (97.2%) and mantle cell lymphoma (95.4%). Burkitt and CD10+ diffuse large B-cell lymphomas were difficult to distinguish by the algorithm. A similar ambiguity was observed between marginal zone, lymphoplasmacytic, and CD10- diffuse large B-cell lymphomas. The specificity of the approach exceeded 98% for all entities. The univariate immunophenotypic library and the multivariate expert-independent diagnostic algorithm might contribute to increased reproducibility of future diagnostics in mature B-cell neoplasms.


Blood ◽  
2012 ◽  
Vol 120 (21) ◽  
pp. 936-936
Author(s):  
Bruno Paiva ◽  
Maria-Carmen Montes ◽  
Ramón García-Sanz ◽  
Jennifer Alonso ◽  
Natalia de las Heras ◽  
...  

Abstract Abstract 936 Demonstration of bone marrow (BM) infiltration by lymphoplasmacytic lymphoma is essential to the diagnosis of WM, and a trephine biopsy is considered mandatory for this assessment. Multiparameter flow cytometry (MFC) has demonstrated its clinical relevance in MGUS and myeloma; however, immunophenotypic studies on IgM monoclonal gammopathies are scanty, and focus only in patients with WM. Herein, MFC immunophenotyping was performed on BM samples from 244 patients, including 67 IgM MGUS, 77 smoldering, and 100 symptomatic WM newly diagnosed patients according to the Second International Workshop. A four color panel that systematically allowed the identification of B cells and plasma cells (PC), and their phenotypic characterization for a total of 24 antigens was used. We first analyzed the percentage of B cells and PC in BM and the percentage of light chain restricted cells in both compartments. Our results show a progressive increment of B cells from IgM MGUS to smoldering and symptomatic WM (medians of 2%, 9% and 12%; P<.001), as well of light chain restricted B cells (75%, 96% and 99%; P<.001). In contrast, no differences were found for the percentage of PC (median of 0.3%), but light chain restricted PC progressively increased from IgM MGUS to smoldering and symptomatic WM (70%, 85% and 97%; P<.001). Accordingly, only 1% of IgM MGUS patients showed >10% B cells, in contrast to 34% and 55% of smoldering and symptomatic WM (P<.001). Likewise, only 1% of IgM MGUS patients showed 100% light chain restricted B cells, in contrast to 19% and 40% of smoldering and symptomatic WM (P<.001); similar results being also found using a cutoff of 100% light chain restricted PC. Subsequently, we explored whether the percentages of BM and light chain restricted B cells and PC could predict time to progression (TTP) from smoldering into symptomatic WM, as well as overall survival (OS) in symptomatic WM. In smoldering WM, B cells (>10% vs ≤10%: median TTP of 47m vs 145m; P=.016) and light chain restricted B cells (100% vs <100%: 26m vs 145m; P<.001) but not PC, predicted risk of progression. On the multivariate analysis that included serum M-spike (±3g/dL), BM infiltration (±50% lymphoplasmacytic cells), BM B-cells and light chain restricted B cells (by MFC), only the later retained independent prognostic value (HR: 19.8, P=.001). Upon analyzing factors influencing survival in symptomatic WM patients, cases with >10% B cells showed a trend for inferior OS (P=.080), and significant differences emerged when comparing patients with 100% vs <100% light chain restricted B cells (median OS 44m vs 78m; P=.001). The later marker was independent (HR: 2.6; P=.004) of the International Prognostic Scoring System (HR: 2.2; P=.006). Focusing on the antigenic profiles of B cells and PC, we noted that within the B-cell compartment there was a progressive increment of CD22dim (69%, 92% and 88%; P<.001), CD25+ (61%, 88% and 90%; P<.001) and sIgM+ (88%, 95% and 97%; P=.002) B cells from IgM MGUS to smoldering and symptomatic WM. This underlies that the accumulating light chain restricted clonal B cells show a characteristic Waldenstrom's phenotype (CD22dim/CD25+/IgM+). Of note, a bimodal (from - to +) expression for the B cell memory marker CD27 was found in >50% of WM patients, which raises the possibility that the WM clone may arise, at least in some cases, before antigenic stimulation; subsequent maturation of the clone into PC would explain the typical presence of somatic hypermutations. On the other hand, B-cells from IgM MGUS and WM patients were negative in ≥90% of cases for CD5, CD10, CD11c and CD103, which can be useful to differentiate between WM and other B-NHL. Finally, the antigenic profile of PC in IgM MGUS and WM was similar to that of normal PC, and different from myeloma PC by consistently showing a CD27+ and CD56- phenotype, in addition to sIgM+ expression in ≥87% of all cases. Similarly to B-cells, we also noted that within the PC compartment there was a progressive increment of CD19+, CD45+ and sIgM+ CD20+ PC from IgM MGUS to smoldering and symptomatic WM. This underlies that this transition is asssociated with an accumulation of light chain restricted clonal PC displaying an immature/plasmablastic phenotype. In summary, our results highlight the potential value of MFC immunophenotyping for the characterization of the Waldenström's clone, as well as for the differential diagnosis, risk of progression and survival in WM. Disclosures: No relevant conflicts of interest to declare.


Blood ◽  
2015 ◽  
Vol 126 (23) ◽  
pp. 2662-2662 ◽  
Author(s):  
Laurent Miguet ◽  
Luc Fornecker ◽  
Marie Wyrwas ◽  
Sarah Cianferani ◽  
Raoul Herbrecht ◽  
...  

Abstract Introduction Diagnosis of mature B-cells proliferations, especially those involving the spleen, do not always falls into any of the WHO types of B-cell neoplasms using standart diagnosis tools. This situation in notably encountered in the case of the differential diagnosis of marginal zone lymphoma (MZL), atypical chronic lymphocytic leukemia (aCLL), mantle cell lymphoma (MCL), and lymphoplasmacytic lymphoma (LPL), mostly due to the lack of immunological positive markers. In order to find new markers to discriminate between these different malignancies, we have previously developed a proteomic strategy based on the analyses of plasma membrane microparticles and proposed two new specific markers: CD148 and CD1801,2 for MCL and MZL respectively. The simultaneous use of these two markers, together with the CD200 that is positive in most cases of CLL and negative in MCL could be of great interest to better assess the differential diagnosis. Methods Flow cytometry analyses have been realized in Nancy and Strasbourg hospitals by combining these three markers: CD148 (Clone 143-41 FITC); CD180 (Clone G28.8 PE) and CD200 (Clone OX104 APC). Expression profile of these proteins was established on a well characterized set of patients (N=287): CLL with a Matutes score > 3 (N=81); MCL harboring t(11;14) translocation or CCND1 overexpression (N=44); LPL (N=58) classified following cytological morphology, IgM peak and the positivity of CD38 and/or Myd88 mutation, MZL (N=84), displaying a CD5- CD23- immunophenotype associated to a splenomegaly and 20 controls. For each group the mean of fluorescence intensity and Standard Error have been determined. Results MCL exhibited a strong expression of CD148 combined with a weak expression of CD180 and CD200. A weak expression of CD148 and CD180 coupled to a strong expression of CD200 was typical of the CLL group and a weak expression of CD148 and CD200 coupled to a strong expression of CD180 was observed in the MZL group. A moderate expression of these three markers was observed in the LPL group. A threshold corresponding to MFI +/- 4 standard error was then calculated for each group, and patients were categorized following the expression profile of these 3 markers (see figures). In this cohort, the above described profiles correctly identified MCL cases with a specificity of 92% and a sensitivity of 64%, aCLL cases with a specificity of 100% and a sensitivity of 47%, LPL cases with a specificity of 90% and a sensitivity of 54% and MZL cases with a specificity of 99% and a sensitivity of 60%. Conclusion These results strongly suggest that the incorporation of these three markers CD148 CD180 and CD200 in addition of the routinely used flow cytometry panel can be helpful in a number of cases for which the diagnosis is difficult. References: 1) Miguet et al leukemia 2013 2) Miguet et al journal of proteome research 2009 Figure 1. Figure 1. Disclosures No relevant conflicts of interest to declare.


Blood ◽  
2013 ◽  
Vol 122 (21) ◽  
pp. 4150-4150 ◽  
Author(s):  
Thomas Köhnke ◽  
Veronika K Wittmann ◽  
Daniela Sauter ◽  
Veit Bücklein ◽  
Zlatana Pasalic ◽  
...  

Abstract Background Immunophenotyping is essential for the diagnosis of chronic lymphocytic leukemia (CLL). The scoring proposed in the modified Matutes score has been the basis of diagnosis for the past 15 years and is defined by strong expression of CD5 and CD23, low or absent expression of CD79b, sIgM and FMC7. However, some markers within the current score such as sIgM display a high variability in staining patterns and thus the interpretation of expression intensity is not easily reproducible. Furthermore, the newly identified marker CD200 is not included in the current score in spite of its highly informative value in the differential diagnosis of B-cell disorders. In the study presented here we aimed to improve the current score through the addition of highly informative markers such as CD200 and the omission of sIgM as a less informative, error-prone marker. Methods Between February 2011 and May 2013, peripheral blood or bone marrow aspirates of patients with suspected B-cell lymphoproliferative disorders were subjected to evaluation by flow cytometry. Immunophenotyping was performed using a Navios flow cytometer (Beckman Coulter) and samples were stained by monoclonal antibodies targeting the antigens CD45, CD19, CD5, CD10, CD23, CD79b, CD200, FMC7, sIgM, kappa and lambda. Corresponding isotype controls were used. The modified Matutes score was calculated as described previously (Moreau et al., Am J Clin Pathol 1997) with positivity defined as ≥20% positive cells. Mean Fluorescence Intensity (MFI) ratio (MFI sample/MFI isotype) was calculated as a measure of expression intensity. For our new score, optimized cut-offs for positivity vs. negativity (CD5, CD23, CD200, FMC7) and low or absent expression (CD79b) as well as sensitivity and specificity were calculated by receiver operating characteristics (ROC). The final clinical diagnosis was defined as the diagnosis established by the treating physician taking into account clinical symptoms as well as all results from diagnostic procedures, including cytomorphology, flow cytometry, cytogenetics, molecular genetics and immunohistochemistry, if available. In order to perform an internal validation of our proposed score, we divided the patient cohort into an exploratory and a validation cohort by a 2:1 ratio based on the date of receipt of the samples. Result Flow cytometry data of 371 patients with B-cell disorders were available for analysis. 247 patients were assigned to the exploratory cohort and 124 patients were assigned to the validation cohort. 84.2% and 82.1% of patients, respectively, were diagnosed with CLL. In the exploratory cohort, sIgM-expression intensity on CD19+ B-cells (as measured by MFI ratio) was significantly lower in CLL versus non-CLL cases (p=0.001). However, low or absent sIgM-expression displayed poor specificity in distinguishing CLL from non-CLL cases (51,3%; sensitivity 83,7%). Absent or low CD79b-expression on CD19+ B-cells showed a higher sensitivity and specificity (94.2% and 71.8%, respectively). Positivity for CD200 as well as lack of FMC7-expression showed high diagnostic value (sensitivity and specificity all above 80%). Interestingly, positivity for CD5 on CD19+ B-cells did not have a strong diagnostic value (sensitivity and specificity 69.7% and 76.9%, respectively), but double positivity for CD5 and CD23 on CD19+ B-cells showed higher sensitivity and specificity (79.8% and 87.2%, respectively). Therefore, CD200+, CD23+/CD5+, FMC7- and low or absent CD79b on CD19+ B-cells were included in a new diagnostic score. The resulting score showed comparable sensitivity (97.1% for our score versus 98.6% for the Matutes score, McNemar’s test p=0.38), but markedly increased specificity (87.2% versus 53.8%, p<0.001). These results were confirmed in the internal validation cohort (sensitivity 97.0% versus 100%, p=N/A; specificity 86.4% versus 59.1%, p=0.03). Conclusion The data support the use of the improved score for the differential diagnosis of CLL. This novel scoring system exhibits significantly higher specificity while maintaining very high sensitivity and might therefore contribute to less false positive results. Finally, the surface markers contained in the novel score show more consistent staining patterns, which might further improve reproducibility. External validation of the proposed score will be pursued. Disclosures: No relevant conflicts of interest to declare.


Blood ◽  
2007 ◽  
Vol 110 (11) ◽  
pp. 4672-4672
Author(s):  
Giuseppe A. Palumbo ◽  
Giovanna Forgione ◽  
Nunzia Parrinello ◽  
Katia Cardillo ◽  
Annalisa Chiarenza ◽  
...  

Abstract Mantle cell lymphoma (MCL) is a B-cell malignancy that shares many morphologic and immunophenotyping features with B-cell chronic lymphocytic leukemia (B-CLL). However, these two entities are characterized by a very different prognosis and response to therapies. Differential diagnosis is assessed by a panel of monoclonal antibodies (moAbs) to screen the expression of CD5, CD23 and Cyclin D1. Nevertheless, some of these antibodies work better on fresh or frozen tissues; for example, on fixed tissues (expecially by B5) CD5 and Cyclin D1 can give equivocal or even negative results. In addition, it is cumbersome to evaluate Cyclin D1 by flow cytometry. CD200 (previously referred to as OX2) is a membrane glycoprotein, belonging to the immunoglobulin superfamily, expressed on T and B lymphocytes (but not on NK cells) and it might play an immunosuppressive role. Its expression has also been reported on human myeloma plasmacells and B-CLL cells. We investigated the expression of CD200 on fresh neoplastic cells of 90 patients (82 B-CLL and 8 MCL in leukemic phase) by flow cytometry, using a mouse IgG1 anti-human moAb (MRC OX-104, BD Pharmingen, U.S.A.). In our series, CD200 was present on neoplastic cells of all 82 B-CLL (expression on 62–100% of CD5+ cells, mean 96%, standard deviation 7%). On the contrary, in MCL patients (Cyclin D1+ and/or t(11;14) FISH+) CD200 was positive in less than 20% of CD5+ cells in 3 subjects (4–18%) and totally absent in the remaining 5 (two-sided Fisher’s exact test p<0.0001). To further extend the study, we examined CD200 by immunohistochemistry on paraffin-embedded formalin-fixed lympoid tissues and Lowy-fixed bone marrow (BM) trephine biopsies from 18 B-CLL (12 lymphnodes and 7 BM samples) and 19 MCL (11 lymphoid tissues and 9 BM) patients, using a goat anti-human IgG affinity-purified polyclonal antibody (R&D Systems, U.S.A.). Again, all B-CLL neoplastic cells were positive for CD200 both in lymphnodes and in BM while all MCL cells were negative, both in lympoid tissues and in BM (two-sided Fisher’s exact test p<0.0001). Notably, in all MCL CD200-negative lympoid tissue biopsies, it was possible to observe CD200+ residual dendritic cells (useful as immunohistochemistry reaction positive control). Moreover, results were consistent in old archival samples and even in non-perfectly prepared tissues referred to our Center. We would propose to add CD200 in flow cytometry and immunohistochemistry routine panels, as it can be of diagnostic utility in distinguishing between MCL and B-CLL, in particular in mantle cell leukemia patients. Finally, these data have to be considered in view of the proposed targeted therapy with anti-CD200, to eventually exclude MCL patients.


2013 ◽  
pp. n/a-n/a ◽  
Author(s):  
Alex F. Sandes ◽  
Maria de Lourdes Chauffaille ◽  
Cláudia Regina M.C. Oliveira ◽  
Yumi Maekawa ◽  
Nair Tamashiro ◽  
...  

2000 ◽  
Vol 64 (4) ◽  
pp. 275-281 ◽  
Author(s):  
Giovanni D'Arena ◽  
Pellegrino Musto ◽  
Nicola Cascavilla ◽  
Matteo Dell'Olio ◽  
Nicola Di Renzo ◽  
...  

2014 ◽  
Vol 142 (3-4) ◽  
pp. 197-203 ◽  
Author(s):  
Tijana Dragovic-Ivancevic ◽  
Nada Kraguljac-Kurtovic ◽  
Vesna Knezevic ◽  
Andrija Bogdanovic ◽  
Biljana Mihaljevic ◽  
...  

Introduction. Accurate diagnosis of chronic lymphocytic leukemia (CLL) acquires immunophenotyping by flow cytometry in order to facilitate differential diagnosis between CLL and other mature B-cell neoplasms (MBCN). Objective. The aim of this study was to define immunological profile of CLL cells. Methods. Immunophenotyping by flow cytometry was performed on peripheral blood specimens at diagnosis in the group of 211 patients with de novo MBCN. Results. Absolute count of B-cells was significantly increased in all MBCN patients comparing to healthy control group (p<0.05). B-cell monoclonality was detected in 96% of all MBCN patients, by using surface immunoglobulin (sIg) light chain restriction. B-cell antigens, CD19, CD20, CD22, were expressed with very high frequency in CLL and other MBCN. In comparison with other MBCN, in CLL group, the frequency of expression was higher for CD5 and CD23 (p<0.0001), though lower for FMC7 antigen (p<0.0001). CLL patients were characterized by lower expression patterns of CD20, CD22, CD79b, and sIg (p<0.0001) as well as higher expression pattern of CD5 antigen (p<0.05). Correlation between the final diagnosis of MBCN and values of CLL scoring system showed that the majority of CLL patients (97%) had higher values (5 or 4) whereas the majority of other MBCN patients (96%) had lower score values (0-3). Conclusion. Our results have shown that characteristic immunophenotype which differentiates CLL from other MBCN is defined by following marker combination - CD19+ CD20+low CD22+low CD5+high CD23+ FMC7- CD79b+low sIg+low. CLL score values of 5 or 4 points are highly suggestive for diagnosis of CLL.


Blood ◽  
2009 ◽  
Vol 114 (22) ◽  
pp. 3940-3940
Author(s):  
Thomas Matthes ◽  
Christiane Ody ◽  
Beat Imhof ◽  
Carmen Donate ◽  
Dominique Cossali ◽  
...  

Abstract Abstract 3940 Poster Board III-876 Introduction Differentiation of naïve B cells into plasma cells or memory cells occurs in the germinal centres (GC) of lymph follicles or alternatively in the marginal zone via a GC- and T cell independent pathway. It is currently assumed that B cell lymphomas correspond to normal B cell differentiation stages, but the precise correlation of several B cell lymphomas to these two pathways remains controversial. We have previously shown that junctional adhesion molecule C (JAM-C) originally identified at the cell-cell border of endothelial cells, constitutes also a marker of B lymphocytes with a tightly regulated expression during B cell differentiation: immature B cells, GC-B cells and plasma cells stain negatively, whereas mature, memory and marginal zone derived B cells stain strongly positive. Here we test the expression of JAM-C on a series of patients with B cell lymphomas. Methods B lymphocytes from the peripheral blood of 158 untreated patients were analyzed using flow cytometry with standard antibody panels (CD5, CD10, CD11c, CD22, CD23, CD25, CD38, CD103, FMC7, sIg). Diagnosis of a B cell lymphoma was established according to WHO guidelines, using additionally RT-PCR, karyotyping, or FISH, if necessary. Expression of JAM-C was studied by flow cytometry with a polyclonal antibody obtained from a rabbit immunized with the soluble JAM-C molecule. Results MCL, HCL and MZBL with a supposed origin in the marginal zone stained mostly positive, whereas CLL and FL with a supposed origin in the germinal centre showed mostly a negative staining. No correlation was found in CLL between JAM-C expression and staining for ZAP70 or CD38. In 12 cases routine work-up was not able to precisely establish a diagnosis of CLL or MZBL, and CLL or MCL. In these cases the presence of JAM-C was considered a strong argument against a GC-origin of the malignant B cells. Addition of JAM-C to antibodies used in the Matutes score increased the sensitivity and specificity of this score for the diagnosis of CLL. Furthermore, it may help differentiating MZBL from LPL which otherwise display overlapping immunophenotypes. Conclusion JAM-C constitutes a new diagnostic marker for the differential diagnosis of B cell lymphomas, and is particularly useful for the distinction between CLL and LPL (negative staining) on the one hand and mantle cell and marginal zone B cell lymphomas (positive staining) on the other hand. Disclosures: No relevant conflicts of interest to declare.


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