In pigs (Sus scrofa), the initial immunoglobulin rearrangement of the κ light chain is replaced by λ before the heavy chains rearrange, and the light chains may rearrange even later. This study investigates whether these developmental differences are reflected in the usage of IGK and IGL genes. We found large differences between peripheral B cells and those developing in the bone marrow, and between B cells in germ-free piglets and conventional pigs. During early B cell development in the bone marrow, more 3' V and 5' J gene segments for both light chains are used. However, in the peripheral naive repertoire, more 5' IGLV and 3' IGLJ genes are used. A similar shift toward the use of more 5' IGKV and 3' IGKJ genes is observed later after antigen exposure in conventional pigs. The expression profile showed that most λ+ B cells are generated earlier, while κ+ B cells develop from late precursors that already contain the λ rearrangement. The initial λ rearrangement is retained in both λ+ and κ+ B lymphocytes, and multiple λ transcripts can be found in individual cells. The overall pool of the IGLV repertoire is therefore much larger and more diversified than for IGKV. The κ repertoire is further restricted to the preferential use of only two major IGKV genes, reflecting the limitation for only two consecutive rearrangements. Tracing of silenced λ transcripts in κ+ B cells further confirmed the unconventional mechanism of differential rearrangements in pigs. Our results underline the diversity of the immune system among mammals.

• Klíčová slova
• B cell development, B cell receptors, Immunoglobulin light chains, Immunoglobulin rearrangement, Lymphocyte differentiation, Porcine immune system,
• MeSH
• B-lymfocyty MeSH
• geny pro imunoglobuliny MeSH
• imunoglobuliny - kappa-řetězce * genetika   MeSH
• imunoglobuliny - lambda-řetězce genetika   MeSH
• lehké řetězce imunoglobulinů * genetika   MeSH
• lymfoidní tkáň MeSH
• prasata MeSH
• savci genetika   MeSH
• zvířata MeSH
• Check Tag
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• Názvy látek
• imunoglobuliny - kappa-řetězce * MeSH
• imunoglobuliny - lambda-řetězce MeSH
• lehké řetězce imunoglobulinů * MeSH

Swine use a reverse order of immunoglobulin chain rearrangement compared to humans and mice, and this altered and modified order should have measurable consequences. Here we perform new and defining experiments with developing and mature B cells, characterizing the B cell populations that do not exist in other species. First, we have finally confirmed that light chains κ and λ are rearranged and expressed on the surface before any heavy chain rearrangements using western-blot. And second, we have analyzed a pool of mature B cells on the single-cell level to demonstrate that many κ+ mature B cells carry λ transcripts. According to these findings, we believe that there may be more groups of mammals; one of which uses a pre-BCR-driven developmental pathway for B cell generation (like mice and humans), the second group uses a pre-BCR-independent one (like swine), and some may be even intermediate.

• Klíčová slova
• B cell development, B cell receptors, Cell differentiation, Immunoglobulin rearrangement, Other animals, Porcine immune system,
• MeSH
• B-lymfocyty MeSH
• geny pro imunoglobuliny * MeSH
• imunoglobuliny - kappa-řetězce * genetika   MeSH
• imunoglobuliny - lambda-řetězce genetika   MeSH
• lehké řetězce imunoglobulinů genetika   MeSH
• lidé MeSH
• myši MeSH
• prasata genetika   MeSH
• savci genetika   MeSH
• zvířata MeSH
• Check Tag
• lidé MeSH
• myši MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• Názvy látek
• imunoglobuliny - kappa-řetězce * MeSH
• imunoglobuliny - lambda-řetězce MeSH
• lehké řetězce imunoglobulinů MeSH

The study aimed to investigate free light chain (FLC) monoclonality in patients with an abnormal free kappa/lambda ratio (FLC ratio). Seventy serum samples with abnormal FLC ratio were examined using an immunoturbidimetry (Binding Site, SPA) and the two different enzyme-linked immunosorbent assays (1. Sebia diagnostic kit; 2. in house methods), the monoclonal or oligoclonal bands of (FLC) by immunofixation electrophoresis (IE) and isoelectric focusing followed by affinity immunoblotting (IEF/AIB). The reference interval was calculated by non-parametric percentile method. 5.7% of samples examined by IE were suspected of monoclonal character of FLCs, but subsequently monoclonality was refuted by more sensitive IEF/AIB method; 7%, resp. 2.9% of samples showed FLC kappa, resp. FLC lambda oligoclonal character of bands. A statistically significant dependence was found between FLC ratio (Sebia) and FLC ratio (SPA), rs = 0.510, p = .001. Kappa statistic evaluated a fair conformity between the FLC ratio (Sebia) and IEF/AIB (kappa = 0.468) and between FLC ratio (in house) and IEF/AIB (kappa = 0.300). The verified reference interval for FLC ratio (Binding Site) is between 0.35 and 2.18. The IEF/AIB is the most sensitive method to discriminate between monoclonal and oligoclonal bands of FLC. The Binding Site and Sebia diagnostic kits do not give consistent results. The Binding Site diagnostic kit provides more results above reference interval of FLC ratios. For routine decision on monoclonality of the FLC ratio (SPA) it is advisable to use a verified reference interval.

• Klíčová slova
• ELISA, Free light chains, immunoturbidimetry, monoclonality, oligoclonal bands,
• MeSH
• dospělí MeSH
• imunoglobuliny - lambda-řetězce analýza   imunologie   MeSH
• isoelektrická fokusace metody   MeSH
• krevní proteiny analýza   MeSH
• lidé středního věku MeSH
• lidé MeSH
• monoklonální protilátky analýza   imunologie   MeSH
• senioři nad 80 let MeSH
• senioři MeSH
• Check Tag
• dospělí MeSH
• lidé středního věku MeSH
• lidé MeSH
• mužské pohlaví MeSH
• senioři nad 80 let MeSH
• senioři MeSH
• ženské pohlaví MeSH
• Publikační typ
• časopisecké články MeSH
• srovnávací studie MeSH
• Názvy látek
• imunoglobuliny - lambda-řetězce MeSH
• krevní proteiny MeSH
• monoklonální protilátky MeSH

Developmental pathways for B cell lymphogenesis are sufficiently known only in mice and humans. However, both of these species rearrange immunoglobulin heavy chains (IgH) before light chains (IgL) while IgL precedes IgH rearrangement in swine. We demonstrate here that this reversed order of rearrangements have some concealed consequences: (1) we confirmed that although IgLκ rearrangement is initial, most IgLλ+ B cells are generated earlier and before IgH rearrangements, while most IgLκ+ B cells later and after IgH rearrangements, (2) the second IgLκ rearrangement can occur after IgLλ rearrangement, (3) early formed B cells bear only single in-frame IgH rearrangements, (4) many IgLκ+ B cells carry IgLλ rearrangements that can be productive and occurring on both alleles in one cell, and (5) although VpreB and λ5 genes are present in swine, they are preferentially expressed in non-B cells. In summary, our findings reveal that swine use an alternative B cell developmental pathway as compared to mice and humans.

• Klíčová slova
• B cell development, B cell receptors, Cell differentiation, Immunoglobulins, Other animals, Porcine immune system,
• MeSH
• B-lymfocyty fyziologie   MeSH
• buněčná diferenciace MeSH
• genová přestavba B-lymfocytů MeSH
• imunoglobuliny - kappa-řetězce genetika   MeSH
• imunoglobuliny - lambda-řetězce genetika   MeSH
• kultivované buňky MeSH
• lidé MeSH
• myši MeSH
• prasata imunologie   MeSH
• receptory antigenů B-buněk genetika   MeSH
• těžké řetězce imunoglobulinů genetika   MeSH
• transkriptom MeSH
• zvířata MeSH
• Check Tag
• lidé MeSH
• myši MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• srovnávací studie MeSH
• Názvy látek
• imunoglobuliny - kappa-řetězce MeSH
• imunoglobuliny - lambda-řetězce MeSH
• receptory antigenů B-buněk MeSH
• těžké řetězce imunoglobulinů MeSH

• Klíčová slova
• array CGH, chronic lymphocytic leukaemia, deletion, lambda light chain, loss of 22q11,
• MeSH
• antigeny nádorové genetika   MeSH
• chromozomální delece * MeSH
• chronická lymfatická leukemie diagnóza   genetika   MeSH
• genetické lokusy * MeSH
• imunoglobuliny - lambda-řetězce genetika   MeSH
• lehké řetězce imunoglobulinů genetika   MeSH
• lidé MeSH
• lidské chromozomy, pár 22 * MeSH
• nádorové biomarkery MeSH
• represorové proteiny genetika   MeSH
• srovnávací genomová hybridizace MeSH
• Check Tag
• lidé MeSH
• Publikační typ
• dopisy MeSH
• práce podpořená grantem MeSH
• Názvy látek
• antigeny nádorové MeSH
• imunoglobuliny - lambda-řetězce MeSH
• lehké řetězce imunoglobulinů MeSH
• nádorové biomarkery MeSH
• PRAME protein, human MeSH Prohlížeč
• represorové proteiny MeSH
• ZNF280B protein, human MeSH Prohlížeč

IgG kappa and IgG lambda concentrations were quantified in 96 paired CSF and sera using Hevylite™ antibodies in an in-house developed sandwich ELISA method. In 56 of these samples, the results were compared with a qualitative isoelectric focusing/affinity-mediated immunoblotting assay for oligoclonal IgG kappa and IgG lambda. Normal IgG kappa/lambda ratio in the CSF was the same as in serum. In 19/33 patients with intrathecal oligoclonal IgG synthesis, skewed IgG kappa/lambda ratio was observed (increased in 16 and decreased in 3 cases). The analysis of light chain composition of intrathecally synthesised immunoglobulins could contribute to our understanding of intrathecal humoral immune response, although its diagnostic utility is limited.

• Klíčová slova
• ELISA, cerebrospinal fluid, immunoglobulins, isoectric focusing, kappa/lambda ratio,
• MeSH
• ELISA metody   MeSH
• imunoglobulin G mozkomíšní mok   imunologie   MeSH
• imunoglobuliny - kappa-řetězce mozkomíšní mok   imunologie   MeSH
• imunoglobuliny - lambda-řetězce mozkomíšní mok   imunologie   MeSH
• lidé MeSH
• Check Tag
• lidé MeSH
• Publikační typ
• časopisecké články MeSH
• Názvy látek
• imunoglobulin G MeSH
• imunoglobuliny - kappa-řetězce MeSH
• imunoglobuliny - lambda-řetězce MeSH

OBJECTIVES: To assess the timelines of serum free light chain (sFLC) concentrations and the kappa/lambda light chain (K/L) ratio in heart transplant (HTX) recipients. To analyze the performance of serum protein electrophoresis (SPE), serum immunofixation (sIFE) and sFLC measurements for gammopathy detection following a HTX. METHODS: A total of 96 patients who underwent a HTX were analyzed during a two-year follow-up period. The relevant clinical data were obtained from patient medical records. SPE, sIFE and sFLC methods were used for the detection of free light chain and intact immunoglobulin gammopathies at 4 time points after HTX. RESULTS: A statistically significant decrease in sFLC K and L (a decrease of 39.1% and 27.6%, respectively, when compared to pretransplant values) was found 9months after the HTX (p<0.001, Friedman test). We detected SPE or sIFE abnormalities in 23 (8.4%) samples, and sFLC K/L ratio abnormalities in 34 (12.4%) samples. All of the K/L ratio abnormalities had normal SPE/sIFE values, and 19% of the findings were persistent. CONCLUSIONS: A significant and consistent dynamics in the sFLC concentration was found in the HTX patients during a 2-year follow-up period, which reflected changes in the immunosuppressant dosage. A remarkable number of monoclonal and polyclonal gammopathies was identified with some persistent abnormalities, using the SPE/sIFE and sFLC methods. Some of the detected abnormalities, which might possess a higher risk for PTLD if interpreted according to common practice in nonTX patients can only be detected by sFLC methods.

• Klíčová slova
• Electrophoresis, Free light chains, Heart transplantation, Immunofixation, Posttransplant lymphoproliferation,
• MeSH
• dospělí MeSH
• imunoglobuliny - kappa-řetězce krev   MeSH
• imunoglobuliny - lambda-řetězce krev   MeSH
• lidé středního věku MeSH
• lidé MeSH
• následné studie MeSH
• senioři MeSH
• transplantace srdce * MeSH
• Check Tag
• dospělí MeSH
• lidé středního věku MeSH
• lidé MeSH
• mužské pohlaví MeSH
• senioři MeSH
• ženské pohlaví MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• Názvy látek
• imunoglobuliny - kappa-řetězce MeSH
• imunoglobuliny - lambda-řetězce MeSH

Randall-type heavy chain deposition disease (HCDD) is a rare disorder characterized by tissue deposition of a truncated monoclonal immunoglobulin heavy chain lacking the first constant domain. Pathophysiological mechanisms are unclear and management remains to be defined. Here we retrospectively studied 15 patients with biopsy-proven HCDD of whom 14 presented with stage 3 or higher chronic kidney disease, with nephrotic syndrome in 9. Renal lesions were characterized by nodular glomerulosclerosis, with linear peritubular and glomerular deposits of γ-heavy chain in 12 patients or α-heavy chain in 3 patients, without concurrent light chain staining. Only 2 patients had symptomatic myeloma. By serum protein electrophoresis/immunofixation, 13 patients had detectable monoclonal gammopathy. However, none of these techniques allowed detection of the nephrotoxic truncated heavy chain, which was achieved by immunoblot and/or bone marrow heavy chain sequencing in 14 of 15 patients. Serum-free kappa to lambda light chain ratio was abnormal in 11 of 11 patients so examined. Immunofluorescence studies of bone marrow plasma cells showed coexpression of the pathogenic heavy chain with light chain matching the abnormal serum-free light chain in all 3 tested patients. Heavy chain sequencing showed first constant domain deletion in 11 of 11 patients, with high isoelectric point values of the variable domain in 10 of 11 patients. All patients received chemotherapy, including bortezomib in 10 cases. Renal parameters improved in 11 patients who achieved a hematological response, as assessed by normalization of the free light chain ratio in 8 cases. Tissue deposition in HCDD relates to physicochemical peculiarities of both variable and constant heavy chain domains. Early diagnosis and treatment with bortezomib-based combinations appear important to preserve renal prognosis. Thus, monitoring of serum-free light chain is an indirect but useful method to evaluate the hematological response.

• Klíčová slova
• free light chains, glomerulosclerosis, immunoglobulin heavy chains, monoclonal gammopathy,
• MeSH
• biopsie MeSH
• bortezomib terapeutické užití   MeSH
• chronická renální insuficience farmakoterapie   imunologie   patologie   MeSH
• fluorescenční protilátková technika MeSH
• glomerulonefritida farmakoterapie   imunologie   patologie   MeSH
• imunoglobuliny - alfa-řetězce analýza   MeSH
• imunoglobuliny - gama-řetězce analýza   genetika   MeSH
• imunoglobuliny - kappa-řetězce analýza   MeSH
• imunoglobuliny - lambda-řetězce analýza   MeSH
• kombinovaná farmakoterapie MeSH
• ledviny účinky léků   imunologie   patologie   MeSH
• lidé středního věku MeSH
• lidé MeSH
• nefrotický syndrom farmakoterapie   imunologie   patologie   MeSH
• nemoc z těžkých řetězců farmakoterapie   genetika   imunologie   patologie   MeSH
• nemoci ledvin farmakoterapie   imunologie   patologie   MeSH
• paraproteinemie farmakoterapie   imunologie   MeSH
• polymerázová řetězová reakce MeSH
• retrospektivní studie MeSH
• senioři nad 80 let MeSH
• senioři MeSH
• výsledek terapie MeSH
• Check Tag
• lidé středního věku MeSH
• lidé MeSH
• mužské pohlaví MeSH
• senioři nad 80 let MeSH
• senioři MeSH
• ženské pohlaví MeSH
• Publikační typ
• časopisecké články MeSH
• multicentrická studie MeSH
• práce podpořená grantem MeSH
• Geografické názvy
• Francie MeSH
• Názvy látek
• bortezomib MeSH
• CHA heavy chain disease protein, human MeSH Prohlížeč
• heavy chain disease proteins, human MeSH Prohlížeč
• imunoglobuliny - alfa-řetězce MeSH
• imunoglobuliny - gama-řetězce MeSH
• imunoglobuliny - kappa-řetězce MeSH
• imunoglobuliny - lambda-řetězce MeSH

OBJECTIVES: We aimed to compare various methods for free light chain (fLC) quantitation in cerebrospinal fluid (CSF) and serum and to determine whether quantitative CSF measurements could reliably predict intrathecal fLC synthesis. In addition, we wished to determine the relationship between free kappa and free lambda light chain concentrations in CSF and serum in various disease groups. METHODS: We analysed 166 paired CSF and serum samples by at least one of the following methods: turbidimetry (Freelite™, SPAPLUS), nephelometry (N Latex FLC™, BN ProSpec), and two different (commercially available and in-house developed) sandwich ELISAs. The results were compared with oligoclonal fLC detected by affinity-mediated immunoblotting after isoelectric focusing. RESULTS: Although the correlations between quantitative methods were good, both proportional and systematic differences were discerned. However, no major differences were observed in the prediction of positive oligoclonal fLC test. Surprisingly, CSF free kappa/free lambda light chain ratios were lower than those in serum in about 75% of samples with negative oligoclonal fLC test. In about a half of patients with multiple sclerosis and clinically isolated syndrome, profoundly increased free kappa/free lambda light chain ratios were found in the CSF. CONCLUSIONS: Our results show that using appropriate method-specific cut-offs, different methods of CSF fLC quantitation can be used for the prediction of intrathecal fLC synthesis. The reason for unexpectedly low free kappa/free lambda light chain ratios in normal CSFs remains to be elucidated. Whereas CSF free kappa light chain concentration is increased in most patients with multiple sclerosis and clinically isolated syndrome, CSF free lambda light chain values show large interindividual variability in these patients and should be investigated further for possible immunopathological and prognostic significance.

• MeSH
• demyelinizační nemoci krev   mozkomíšní mok   diagnóza   MeSH
• ELISA přístrojové vybavení   metody   MeSH
• imunoblotting přístrojové vybavení   metody   MeSH
• imunoglobuliny - kappa-řetězce biosyntéza   krev   mozkomíšní mok   MeSH
• imunoglobuliny - lambda-řetězce biosyntéza   krev   mozkomíšní mok   MeSH
• isoelektrická fokusace přístrojové vybavení   metody   MeSH
• lidé MeSH
• nefelometrie a turbidimetrie přístrojové vybavení   metody   MeSH
• odchylka pozorovatele MeSH
• reprodukovatelnost výsledků MeSH
• ROC křivka MeSH
• roztroušená skleróza krev   mozkomíšní mok   diagnóza   MeSH
• studie případů a kontrol MeSH
• Check Tag
• lidé MeSH
• Publikační typ
• časopisecké články MeSH
• srovnávací studie MeSH
• Názvy látek
• imunoglobuliny - kappa-řetězce MeSH
• imunoglobuliny - lambda-řetězce MeSH

INTRODUCTION: Assessment of serum levels of free light chains (FLC-κ and FLC-λ) and recently heavy/light chain pairs of immunoglobulin (HLC-κ and HLC-λ) and their ratio (FLC-r and HLC-r) has significantly enriched traditional algorithm of multiple myeloma (MM) evaluation. The aim of the presented study was to assess the relationship of classical prognostic parameters of MM, standard FLC-κ/λ and HLC-κ/λ ratio ((s)FLC-r and (s)HLC-r), modified ratio of "involved/uninvolved" FLC and HLC ((m)FLC-r and (m)HLC-r ), the difference between "involved - uninvolved" FLC and HLC (FLC-dif. and HLC-dif.) to current stratification models of MM based on the result of cytogenetic analysis. PATIENTS AND METHODS: In a group of 97 patients with MM we assessed serum levels of FLC by FreeliteTM method, and we calculated (s)FLC-r, (m)FLC-r and FLC-dif. indices by HevyliteTM method. For cytogenetic analysis we used FICTION (fluorescence immunophenotyping and interphase cytogenetics as a tool for the investigation of neoplasms). For MM stratification we used standard staging systems according to Durie-Salmon (D-S) and International Staging System (ISS) as well as novel stratification systems based on the results of cytogenetic analysis, ie. "Mayo Stratification of Myeloma and Risk-Adapted Therapy" (mSMART) and "Revised International Staging System" (R-ISS). RESULTS: Stratification mSMART and R-ISS has significantly different representation of "standard" or "low-risk" (71, 15.5, 11.3 a 29.9 %), "intermediate risk" (15.5, 53.6, 34 a 33 %) and "high risk" patients (13.4, 30.9, 54.7 a 37.1 %) compared to standard staging systems. mSMART stratification was compared to prognostic factors of MM (Hb, albumin, β(2)-M, creatinine and LDH), and the only significant relationship was found in the case of β(2)-M, R-ISS had relationship only to Hb and creatinine. In the case of D-S staging we found significant relationship of stages 1-3 and substages A and B to the levels of (m)FLC-r, FLC-dif. and (m)HLC-r, ISS had moreover relationship to k HLC-dif. and MIg concentration. Analysis of mSMART stratification showed primarily significant relationship of risk categories 1-3 to (m)FLC-r and (s)HLC-r indices, and R-ISS to (m)HLC-r index and MIg concentration. In both cytogenetics-based stratifications there was a lack of relationship to (s)FLC-r, FLC-dif. and HLC-dif. indices. CONCLUSION: Comparison of the results of standard staging systems according to D-S and ISS with cytogenetics based models mSMART and R-ISS showed different representation of risk groups, and significantly different relationship to classical prognostic factors together with original relationship of sMART stratification to (m)FLC-r and (s)HLC-r, and R-ISS to (m)HLC-r and MIg concentration.

• MeSH
• dospělí MeSH
• imunoglobuliny - lambda-řetězce krev   MeSH
• lehké řetězce imunoglobulinů krev   MeSH
• lidé středního věku MeSH
• lidé MeSH
• mnohočetný myelom krev   patologie   MeSH
• prognóza MeSH
• senioři nad 80 let MeSH
• senioři MeSH
• Check Tag
• dospělí MeSH
• lidé středního věku MeSH
• lidé MeSH
• mužské pohlaví MeSH
• senioři nad 80 let MeSH
• senioři MeSH
• ženské pohlaví MeSH
• Publikační typ
• časopisecké články MeSH
• Názvy látek
• imunoglobuliny - lambda-řetězce MeSH
• lehké řetězce imunoglobulinů MeSH