Serum Of Multiple Myeloma Patients Research Articles

Multiple myeloma associated with sialyl salivary-type amylase

BackgroundThere have been several reports describing a notable hyperamylasaemia in patients with multiple myeloma. Such amylase-producing myelomas have been mainly described in the context of concomitant salivary-type hyperamylasaemia, with sialyl salivary-type amylase identified in a portion of those cases. We investigated the incidence of the production of sialyl salivary-type amylase in serum of multiple myeloma patients. MethodsEleven patients (6 male and 5 female) who had been diagnosed as having multiple myeloma were enrolled in this study. Sialyl salivary-type amylase was detected by isoamylase electrophoresis and HPLC analysis, and identified by detecting either abnormal neuraminidase-sensitive band through isoamylase electrophoresis or abnormal extra-elution peak of amylase by means of HPLC analysis. ResultsSialyl salivary-type amylase was detected in 7 out of 11 (63.6%) patients. Median total amylase activity was 154 U/l (range 109–43020). Isoamylase electrophoretic patterns of patients' serum were normal in 5 patients (71.4%) out of 7 patients and salivary-dominant in 2 (50.0%) out of 4 patients. ConclusionsWe consider that there is no significant relationship between total serum amylase level and amylase isoenzyme pattern in the incidence of production of sialyl salivary-type amylase with multiple myeloma.

Serum Pleiotrophin Is a New Multiple Myeloma Tumor Marker That Also Predicts Clinical Status.

Pleiotrophin (PTN), an 18 kD heparin-binding protein, has been found to be expressed in some solid tumors whereas it has not been previously evaluated in hematologic malignancies. Constitutive expression of PTN promotes tumor expansion and may increase metastatic potential. This protein binds to syndecan-1 (CD138) and stimulates angiogenesis. Its role in multiple myeloma (MM), thus far, has not been evaluated. Recently, we showed that PTN was strongly expressed in MM cell lines and malignant plasma cells from fresh bone marrow samples but not in normal control bone marrow specimens. Since PTN has been shown to be elevated in the serum of some solid tumor patients, we determined whether PTN may be elevated in the serum of MM patients, and correlated these levels with the patients' disease status. By using a highly sensitive and specific enzyme-liked immunosorbent assay (ELISA) for PTN, we tested 270 different serum samples [MM (n=194), MGUS (n=18) and age-matched healthy control subjects (n=58)]. Serum samples from MM patients were obtained at the time of diagnosis as well as during the course of their disease. Serum PTN levels were higher in MM patients than in the control subjects (median=1.44 ng/ml vs 0.42 ng/ml, p <0.0001). In addition, among patients with MGUS and smoldering myeloma, serum PTN was also higher than in the controls (median=1.14 ng/ml vs 0.42 ng/ml, p <0.0001) but lower than among patients with active MM (p <0.0001). Patients with untreated MM showed serum PTN levels similar to those patients who received prior treatment. Among previously treated patients, the group with progressive disease at the time of evaluating their PTN level had higher serum concentrations of this protein than patients with responsive or stable disease (median=1.73 ng/ml vs 1.16 ng/ml, p <0.0001). Evaluation of patients who had a change in disease status showed that the serum PTN level increased at the time of progression when compared with their baseline level (median=1.76 ng/ml vs 1.01 ng/ml, p=0.009). Conversely, patients who responded to anti-MM therapy exhibited significant decreases of PTN as compared with their pre-treatment PTN values (median=0.95 ng/ml vs 1.90 ng/ml, p=0.007). These results suggest that serum PTN may be a new tumor marker for MM, and monitoring of this protein may be useful to follow the responsiveness to therapy and disease status of MM patients. We are now undertaking studies to evaluate PTN as a prognostic marker in a large cohort of newly diagnosed MM patients.

Myeloma paraprotein epitope identification using a phage library of sterically constrained peptides

6626 Background: Chronic antigenic stimulation has been hypothesized to be a causative factor in rare cases of multiple myeloma (MM). If true, one might identify relevant antigens by using phage display technology and biopanning to analyze epitopes of m-proteins in sera of MM patients (pts). This method obviates the need to prospectively identify candidate antigens. Methods: The Ig fractions of 17 pts with IgG paraproteins were purified on Protein G beads. The bead-bound m-proteins were then used to biopan a T7 library of random heptapeptides sterically constrained by intramolecular disulfide bridges (Gift of G. Froman). Phage selected via 3–5 rounds of biopanning were cloned by plaque isolation. Binding specificity was confirmed by blotting and chemiluminescent detection. The cloned phage inserts’ DNA was sequenced and the amino acid sequences of each insert deduced. Proteins containing homologies with these peptides were found by short-peptide BLAST search. To test whether the identified epitopes are common among MM pts, the phage clones were screened against IgG m-proteins from sera of 70 additional pts. Results: Phage clones were identified which reacted specifically with 3 of the 17 pt m-proteins in the panel. There were 7 selected clones in total, as multiple clones were found for two pts. None of the peptide sequences displayed strong homology with known human autoantigens. However, the peptide inserts of the isolated clones exhibited strong homologies with proteins of pathogens including Mycoplasma sp., Fusobacterium sp., influenza A, and Candida sp., some of which are antigens ubiquitously present in the environment. None of the Ig from 70 other sera cross-reacted with any of the selected phage. Conclusions: We have shown at least some MM paraproteins bind epitopes expressed by known human pathogens, supporting the possible role of chronic antigenic stimulation in myeloma pathogenesis. These findings are consistent with a previously published report (Scand. J. Immunol. 57:583;2003). In addition, the lack of any cross-reacting Igs from the large panel of sera makes it statistically unlikely that any of these pathogens, even if contributory to individual cases, are commonly causative. No significant financial relationships to disclose.

Serum Soluble Syndecan-1 Levels Correlate with Erythropoietin and Inversely with Hemoglobin Levels in Untreated Anaemic Myeloma Patients: Possible Role of Syndecan-1 in the Mechanisms of Chronic Anaemia.

Impaired erythropoiesis with inappropriate erythropoietin (EPO) production is seen in about 40% of symptomatic multiple myeloma (MM) patients at diagnosis and worsens during terminal refractory disease. Up-regulation of proinflammatory and proapoptotic factors, namely TNF-alpha, INF-gamma, TGF-beta1, FAS-Ligand and TRAIL are involved in the pathogenesis of defective erythroid maturation. The involvement of these factors in the progression of MM has been studied and no correlation was found for TNF-alpha, INF-gamma and TGF-beta1, while it was shown that the up-regulation of FAS-L/TRAIL in MM cells contributes to MM aggressiveness. Increased soluble syndecan-1 (s-synd-1) serum levels have been shown to constitute an adverse prognostic factor for survival in MM. Furthermore at least 3 studies, including ours, also found a negative correlation of s-synd-1 levels with hemoglobin. The purpose of this study was to investigate the correlations between s-synd-1, hemoglobin (Hb), endogenous EPO levels and other parameters of disease in anaemic MM patients before any treatment including rHu-EPO administration. 41 MM patients (19 males and 22 females) with variable degree of anaemia (Hb <13.5 g/dl for men and <12.5 g/dlfor women) were studied. Median Hb was 10.5 g/dl (range 6.9–13.5); 32% had Hb below 10 g/dl. Median age was 68y. 7 patients had Durie and Salmon stage I, 21 stage II and 13 stage III. Serum EPO levels were determined by RIA and s-synd by ELISA (Diaclone Research, Besancon, France) in frozen sera of MM patients and 24 healthy individuals. Normal serum EPO levels range from 4.9 to 52 mU/ml in our laboratory. Median s-synd-1 was 36 ng/ml (9–75) in healthy individuals.The median value of EPO levels in MM patients was 40.1 mU/ml (7.2–313) and of s-synd-1 160 ng/ml (26–3500). Serum EPO levels correlated as expected with Hb (Spearman's rho -0.223, p=0.14), creatinin (Spearman's rho -0.32, p=0.04) but also with s-synd-1 (Spearman's rho 0.33, p=0.03). Serum EPO did not correlate either with parameters reflecting disease mass such as beta2microglobulin (Spearman's rho −0.19, p=0.90) and percentage of bone marrow infiltration (Spearman's rho 0.118, p=0.48) or CRP (Spearman's rho −0.189, p=0.29) that reflects proinflammatory cytokines' activity. Our results in addition to the fact that s-synd-1 levels inversely correlate with Hb levels, suggest that s-synd-1 may be involved in the mechanisms leading to chronic anemia in multiple myeloma. Further studies are needed to confirm these findings.

The Proteomic Analysis of Sera for the Discovery of Biomarkers for Multiple Myeloma.

Multiple Myeloma (MM) fits in the group of plasm cell disorders characterized by neoplastic proliferation of single clone of plasma cell engaged in the production of a monoclonal immunoglobulin. The cause of MM is unclear, but a genetic predisposition may play a role and cytogenetic abnormalities are considered to be the most important prognostic factor. The proteomic analysis of MM emerges as a key complement to gene expression profiling, primarily because regulation of protein expression can buffer the magnitude of changes occurring at the gene transcription level. In the present study, to identify biomarkers of initial diagnosis, detection of relapse, monitoring for minimal residual disease and prognostic marker in MM by a lesser invasive method, serum proteins reflecting alteration in their proteomes were analyzed. We compared two-dimensional electrophoresis patterns of human sera of twelve patients with multiple myeloma with that of normal twelve subjects. The differentially expressed spots were identified by matrix-assisted laser desorption/ionization-time-of flight (MALDI-TOF) and electrospray ionization quadupole time of flight (ESI-Q-TOF) mass spectrometry. We identified several proteins altered in sera of MM patients. These results suggest that these proteins can be used as a lesser invasive diagnostic, monitoring and prognositic biomarkers of MM if further studies were done.

Change in relative mobility of M protein following neuraminidase treatment in patients with multiple myeloma and monoclonal gammopathy of undetermined significance.

The aim of this study was to clarify the relationship between the relative mobility of M protein in various disease states using cellulose acetate membrane electrophoresis. To examine the carbohydrate chain of the M protein, sera from patients with multiple myeloma (MM), various cancers, and benign disease were treated with neuraminidase. The relative mobility in benign disease and MM patient sera following neuraminidase treatment varied among individuals, while that of IgG M protein in sera from cancer patients was from 0.2 to 0.3. Thus, the relative mobility in cancer patients was narrower than in those with MM or benign disease.However, after neuraminidase treatment, there was no significant difference between relative mobility in cancer patient's sera and those in other disease patients.

Interleukin-10 Is a Proliferation Factor But Not a Differentiation Factor for Human Myeloma Cells

Because interleukin-10 (IL-10) is a potent differentiation factor of human B cells into mature plasma cells, we investigated its effect on human malignant plasma cells. IL-10 did not induce any differentiation and increase in Ig synthesis in four human IL-6-dependent malignant plasma cell lines. However, it stimulated the proliferation of two of four cytokine-dependent cell lines in the absence of IL-6 and IL-10-dependent myeloma cell lines have been obtained. The myeloma cell growth activity of IL-10 was unaffected by anti-IL-6 and anti-IL-6R antibodies. Similarly, IL-10 stimulated (P = .001) the proliferation of freshly-explanted myeloma cells in IL-6-deprived cultures of tumor samples from patients with active multiple myeloma (MM) and produced twice as many myeloma cells in these cultures. Again, this cytokine was unable to induce further differentiation (assessed by rate of Ig production) of fresh myeloma cells. A very sensitive enzyme-linked immunosorbent assay (ELISA; 1 pg/mL) only rarely detected IL-10 in the sera of MM patients (3 of 89). On the contrary, serum IL-10 was detected in 60% of patients with plasma cell leukemia (12 of 20). These data show that IL-10 is an IL-6-unrelated growth factor for malignant plasmablastic cells. This cytokine could be involved in the late phase of MM in vivo.

Expression of MUC1 on myeloma cells and induction of HLA-unrestricted CTL against MUC1 from a multiple myeloma patient.

Polymorphic epithelial mucin (MUC1) was detected in myeloma cells and in sera of multiple myeloma patients. HLA-unrestricted CTL that recognize tumor-associated epitopes on MUC1 has been shown to be induced from breast and pancreas cancer patients. To investigate whether such CTL can also be induced from multiple myeloma patients, an allogeneic mixed leukocyte tumor cell culture was performed. PBMCs of a multiple myeloma patient were stimulated by different allogeneic breast carcinoma and myeloma cell lines. The cultured PBMCs were proliferated and a CTL line TN was established. TN exclusively expressed TCR-alpha/beta, CD3, and CD8. TN lysed breast carcinoma and myeloma cell lines but did not lyse K562, which is sensitive to NK cells. The cytotoxicity of TN was inhibited by anti-CD3 Abs but not by anti-HLA Abs. Thus, the TCR-alpha/beta was considered to be involved in the recognition of the target cells but HLA was not. Furthermore, TN lysed transformed mouse fibroblast cells transfected with MUC1 cDNA, suggesting that this CTL line recognizes MUC1 directly. Thus, it is concluded that precursors of HLA-unrestricted and anti-MUC1 reactive CTL could exist in the peripheral blood of multiple myeloma patients and that myeloma cells can express epitopes on MUC1, which can be recognized by the CTL.

Immune complexes in human myeloma and rat immunocytoma.

1. CIC levels of normal rats show characteristic fluctuations; 2. the implantation of Ig producing immunocytoma cells is followed by the sudden increase of CIC and a more expressed fluctuation on significantly higher levels; 3. the increase of CIC level in the early phase of tumor implantation is not the consequence of humoral immune response induced by the Ig produced by the immunocytoma cells, but it may be due to an interaction of tumor specific antigens and cross-reacting natural antibodies; 4. the CIC content in the sera of healthy persons shows a characteristic fluctuation. The results obtained by different assays are seldom parallel pointing to the heterogeneity of CIC; 5. CIC levels in sera of multiple myeloma patients are usually increased and show no correlation with the monoclonal Ig content of the sera.

The identification of the subclasses of human IgG by analytical isotachophoresis

A new method is described for demonstrating the individual subclasses of human IgG immunoglobulins by means of analytical isotachophoresis. Individual subclasses of IgG purified from sera of multiple myeloma patients have distinct mobility patterns. When a mixture with purified IgG for each of the 4 subclasses is analyzed, a characteristic mobility pattern with 4 unique peaks is obtained. An IgG of unknown subclass can be identified by the superimposition of the peak for the unknown upon one of the 4 well characterized peaks. The method using microgram quantities of purified protein in a 1.0-μl volume can be performed easily in 15 min.

AGAMMAGLOBULINEMIA IN ADULTS

Case Reports1 March 1957AGAMMAGLOBULINEMIA IN ADULTSJ. W. SAVACOOL, M.D., F.A.C.P., RAY P. LANDES, M.D.J. W. SAVACOOL, M.D., F.A.C.P.Search for more papers by this author, RAY P. LANDES, M.D.Search for more papers by this authorAuthor, Article, and Disclosure Informationhttps://doi.org/10.7326/0003-4819-46-3-628 SectionsAboutPDF ToolsAdd to favoritesDownload CitationsTrack CitationsPermissions ShareFacebookTwitterLinkedInRedditEmail ExcerptThe purpose of this report is to call attention to another case of agammaglobulinemia in an adult, to emphasize a few of the features of this interesting process, and to add to the accumulating data with respect to its association with various relatively common pathologic processes.CASE REPORTA 38 year old white male clerk was admitted to Grand View Hospital May 29, 1954, with an 18 month history of recurrent episodes of fever, chills, sweats, coughs, expectoration, bilateral anterior chest pain and weakness.As a child he had apparently had good health, having had only measles. At about the...Bibliography1. SanfordFavourTribeman JPCBMS: Absence of serum gamma globulin in an adult, New England J. Med. 250: 1027-1029 (June 17) 1954. CrossrefMedlineGoogle Scholar2. Bruton OC: Agammaglobulinemia, Pediatrics 9: 722-728 (June) 1952. MedlineGoogle Scholar3. PrasadKoza ASDW: Agammaglobulinemia, Ann. Int. Med. 41: 629-639 (Sept.) 1954. LinkGoogle Scholar4. ZinnemanHallHeller HHWHBI: Acquired agammaglobulinemia, J. A. M. A. 156: 1390-1393 (Dec. 11) 1954. CrossrefMedlineGoogle Scholar5. RohmBehnkeBond RJRHWH: Acquired agammaglobulinemia and hypersplenism, J. Lab. and Clin. Med. 44: 918, 1954. Google Scholar6. GrantWallace AHWD: Agammaglobulinemia, Lancet 2: 671-673 (Oct. 2) 1954. CrossrefGoogle Scholar7. WallSaslaw RLS: Adult agammaglobulinemia, Arch. Int. Med. 95: 33-36 (Jan.) 1955. CrossrefGoogle Scholar8. Verschure JD: Gammaglobulinopenia, Nederl. tijdschr. v. geneesk. 98: 2805, 1954. MedlineGoogle Scholar9. CraigGitlinJewett JMDTC: The response of lymph nodes of normal and congenitally agammaglobulinemic children to antigenic stimulation, Am. J. Dis. Child. 88: 626 (Nov.) 1954. Google Scholar10. JanewayCraigDavidsonDowneyGitlinSullivan CAJAMWDJC: Hypergammaglobinemia associated with severe recurrent and chronic nonspecific infection, Am. J. Dis. Child. 88: 388, 1954. Google Scholar11. HamburgerPetermann FMC: Studies on hypoproteinemia. II. Familial idiopathic dysproteinemia: Blood 4: 1085-1108 (Oct.) 1949. CrossrefMedlineGoogle Scholar12. Good RA: Agammaglobulinemia: a provocative experiment of Nature, Bull. Univ. Minnesota Hospitals and Minnesota Med. Foundation 26: 1, 1954. Google Scholar13. YoungWolfson IIWQ: The agammaglobulinemia syndrome in adult men: its differentiation into familial lymphopenic agammaglobulinemia, Clin. Res. Proc. 2: 101, 1954. Google Scholar14. GoodKelley RAVC: Adrenal cortical function in patients with agammaglobulinemia, Proc. Soc. Exper. Biol. and Med. 88: 99-101 (Jan.) 1955. CrossrefMedlineGoogle Scholar15. LaskiSass-KorstakHillman BADA: Cyclic neutropenia and agammaglobulinemia, Am. J. Dis. Child. 88: 820-821, 1954. Google Scholar16. ReinerStern MKG: Electrophoretic studies on the protein distribution in the serum of multiple myeloma patients, Acta hematol. 9: 19-29 (Jan.) 1953. CrossrefMedlineGoogle Scholar17. Brownell EG: Multiple myeloma, Arch. Int. Med. 95: 699-704 (May) 1955. CrossrefGoogle Scholar18. Reimann HA: Periodic disease: report of old cases, new cases and therapy, Arch. Int. Med. 92: 494-506 (Oct.) 1953. CrossrefGoogle Scholar This content is PDF only. To continue reading please click on the PDF icon. Author, Article, and Disclosure InformationAffiliations: Philadelphia, Pennsylvania*Received for publication November 5, 1955.From the Barton Division and the Department of Medicine, Jefferson Medical College Hospital, Philadelphia, and the Grand View Hospital, Sellersville, Pennsylvania.Requests for reprints should be addressed to J. W. Savacool, M.D., 146 West Tulpehocken Street, Philadelphia 44, Pennsylvania. PreviousarticleNextarticle Advertisement FiguresReferencesRelatedDetails Metrics Cited byIdiopathic Acquired Agammaglobulinemia Associated with ThymomaHOST-PARASITE RELATIONSHIPS IN PATIENTS WITH DYSPROTEINEMIAS 1 March 1957Volume 46, Issue 3Page: 628-637KeywordsAntibioticsChild healthFeversHospital medicineIschemiaMeaslesPneumoniaProteinsSweatUpper respiratory tract infections Issue Published: 1 March 1957 PDF downloadLoading ...

Electrophoretic studies on the protein distribution in the serum of multiple myeloma patients.

Electrophoretic studies on the protein distribution in the serum of multiple myeloma patients.