ࡱ> MOJKLY :SbjbjWW "==+]....222FFFF8~L<F+000)))))))$, .*2"*..00.020)FF....)^)"V)@J2)0p4FFB) MULTIPLI MIJELOM S MONOKLONSKIM IMUNOGLOBULINOM D IgD MULTIPLE MYELOMA Danica Matiai1, Dubravka voriaec1, Mileva Trogrli2, Maja Pauro2, Marija Tatkovi3 Klini ki zavod za laboratorijsku dijagnostiku, Medicinski fakultet Sveu iliata u Zagrebu i Klini ki bolni ki centar Zagreb1; Kiapatieva 12, Zagreb Djelatnost za laboratorijsku dijagnostiku2 i Djelatnost za unutraanje bolesti3 Ope bolnice Pula Adresa za dopisivanje: Dr. sc. Danica Matiai, Klini ki zavod za laboratorijsku dijagnostiku, Medicinski fakultet Sveu iliata u Zagrebu i Klini ki bolni ki centar Zagreb1; Kiapatieva 12, Zagreb Tel.:23 88 207; Fax: 23 12 079; E-mail: dmatisic@kbc-zagreb.hr SA}ETAK - Cilj: prikazati laboratorijsku dijagnostiku multiplog mijeloma s monoklonskim imunoglobulinom D (IgD MM). Metode: elektroforeza proteina u serumu na acetat-celulozi; imunonefelometrijsko odreivanje imunoglobulina G, A i M; imunoelektroforeza za definiranje monoklonskog imunoglobulina D (IgD); imunoelektroforeza za dokazivanje monoklonskih slobodnih laganih lanaca imunoglobulina (proteina Bence Jones-BJP) u serumu i imunofiksacija za dokazivanje BJP u mokrai. Rezultati: kod prikazanog bolesnika dokazan je monoklonski IgD tipa lambda kao i BJP tipa lambda u serumu i mokrai. Zaklju ci: IgD MM je rijetka bolest koja je uzrokom samo 2% novodijagnosticiranih slu ajeva multiplog mijeloma, te obi no ima agresivan tijek i loau prognozu. Laboratorijsku dijagnostiku monoklonskog IgD treba provesti prema  Dijagnosti kom algoritmu za definiciju monoklonskog proteina , koji je sastavio Pododbor za laboratorijsku hematologiju i morfologiju Nacionalne grupe za lije enje hematoloakih bolesti. Klju ne rije i: multipli mijelom; IgD; BJP tipa lambda; imunofiksacija UVOD Imunoglobulin D (IgD) je jedan od pet razreda imunoglobulina i ini oko 0,2% serumskih antitijela. Javlja se kao monomer i ima 2 epitopa. Nalazi se u vrlo niskoj koncentraciji u serumu, a njegova uloga joa nije razjaanjena (4,5). IgD su prvi put opisali Rowe i Fahey 1965. godine (1). Primarno je naen na povraini B-limfocita gdje funkcionira kao receptor za antigen. IgD na povraini B-limfocita ima posebne aminokiseline na C-terminalnom kraju kojima se  sidri za membranu. IgD ne ve~e komplement (7). Manjak IgD je defekt humoralnog imuniteta. Slabo je poznata i normalna funkcija IgD, a malo je i klini kih znakova ili simptoma koji prate odsutnost njegove sinteze. Osobe s vrlo niskom koncentracijom IgD ili njegovim nedostatkom ne pokazuju veu sklonost infekciji. Genetska preuredba, koja se dogaa za vrijeme sazrijevanja B- limfocita, rezultira izra~ajem IgM i IgD na povraini zrelog B-limfocita. Stvaranje IgD je stimulirano s IL-4 i IL-10 (6). Multipli mijelom s monoklonskim imunoglobulinom D (IgD MM) javlja se vrlo rijetko, u svega 2% od svih novootkrivenih bolesnika s multiplim mijelomom (3). U veini bolesnika nalazi se hipogamaglobulinemija. Isto tako, oateenje bubrega prisutno je u 67% bolesnika. Osteoliti ka ~ariata nalaze se kod 75% bolesnika s IgD MM. IgD MM esto prate monoklonski slobodni laki lanci imunoglobulina (proteina Bence Jones-BJP), za razliku od IgG i IgA MM gdje je pojavnost BJP od 16%-33%. U veini bolesnika s IgD MM (89%-94%) nalazi se BJP tipa lambda, u usporedbi s IgG ili IgA MM gdje su BJP tipa lambda naeni samo kod 1/3 bolesnika (2). U 44% slu ajeva IgD MM naena je i amiloidoza. Poznato je da pribli~no 50% bolesnika s IgD MM ima popratnu organomegaliju i limfadenopatiju. Iako je odgovor na terapijske tretmane sli an kao i kod ostalih tipova multiplog mijeloma, medijan pre~ivljavanja je mnogo krai kod IgD MM (12 17 mjeseci) (3). U razdoblju od 1992. do 2004. godine u naaem laboratoriju naeno je pet slu ajeva IgD MM od kojih jedan prikazujemo. PRIKAZ BOLESNIKA Muakarac u dobi od 46 godina hospitaliziran je putem hitne slu~be zbog nepodnoaenja napora, ope slabosti te subfebriliteta. Tegobe su po ele nakon respiracijskog infekta. Nakon terapije antibiotikom u injena je osnovna laboratorijska obrada te zbog ubrzane sedimentacije eritrocita i normocitne anemije s po etnom bubre~nom insuficijencijom upuen je u bolnicu. Kod prijama u bolnicu nalazi se ubrzana sedimentacija eritrocita. U perifernoj krvnoj slici naene su rouleaux-formacije eritrocita. Odreeni su ukupni proteini, imunoglobulini G, A i M i u injena je elektroforeza proteina u serumu. Naeni su izrazito poviaeni ukupni proteini, izrazito sni~ene vrijednosti navedenih imunoglobulina, a elektroforezom proteina u serumu naene su dvije monoklonske vrpce. Prema vrijednostima imunoglobulina G, A i M bilo je o ito da monoklonske vrpce ne pripadaju niti jednom od njih, te je zbog sumnje na monoklonski IgD u injena i imunoelektroforeza s antiserumima protiv teakih lanaca delta i vezanih lakih lanaca kapa i lambda tipa kao i BJP tipa kapa i lambda u serumu i u 100 x uguaenoj mokrai. Imunoelektroforezom seruma naeno je da jedna vrpca potje e od monoklonskog IgD tipa lambda, a druga od BJP tipa lambda. U mokrai su takoer naeni BJP tipa lambda (slika1). U injena je takoer i sternalna punkcija gdje je naena poviaena infiltracija plazma stanicama. Rendgenskom obradom kostiju nisu naena osteoliti ka ~ariata. S obzirom na dijagnosti ke kriterije plazmocitoma i ~ivotnu dob zapo eto je lije enje kemoterapijom po protokolu: Vinkristin, Adriamicin, Dexamethason (VAD). Rezultati osnovne laboratorijske obrade kod prijama bolesnika, nakon prvog i petog ciklusa kemoterapije te neposredno prije i poslije autologne transplantacije krvotvornih stanica, prikazani su u tablici 1. Tablica 1. Rezultati laboratorijske obrade Table 1. Laboratory test results Na temelju nalaza u injene osnovne laboratorijske obrade nakon 1. i 5. ciklusa VAD- terapije, mo~e se zaklju iti da se radilo o djelomi nom odgovoru na kemoterapiju. Tri mjeseca nakon zavraene kemoterapije bolesnik je pripremljen za autolognu transplantaciju krvotvornih mati nih stanica. Iako se po u injenim laboratorijskim nalazima radilo o potpunoj remisiji bolesti, za potpunu potvrdu napravljena je i kontrolna sternalna punkcija koja je to i potvrdila (4% plazma stanica u nalazu sternalnog punktata). Rezultati elektroforeze proteina u serumu, kao i imunofiksacije s BJP tipa lambda i kapa u 100 x uguaenoj mokrai poslije autologne transplantacije krvotvornih mati nih stanica prikazani su na slici 2. Slika 1. Elektroforeza proteina u serumu i imunoelektroforeza seruma i 100 x uguaene mokrae Figure 1. Serum protein electrophoresis and immunoelectrophoresis of serum and 100 x concentrated urine Slika 2. Elektroforeza proteina u serumu i imunofiksacija 100 x uguaene mokrae Figure 2. Serum protein electrophoresis and immunofixation of 100 x concentrated urine RASPRAVA IgD MM je vrlo rijetka bolest i javlja se u svega 1 2% svih novootkrivenih slu ajeva multiplih mijeloma (2). Multipli mijelom je bolest starije dobi i veinom se javlja u 60.-im godinama ~ivota, meutim IgD MM se javlja mnogo ranije i zbog vrlo niske koncentracije u serumu kasno se uo ava kao i multipli mijelom lakih lanaca, pa gotovo svi bolesnici budu otkriveni tek u treem stadiju bolesti. Svih pet bolesnika otkrivenih u naaem laboratoriju naeni su u treem stadiju bolesti. Iako se o IgDMM mo~e nai relativno malo objavljenih radova, studije koje su provedene pokazuju da se eae pojavljuje kod muakaraca nego kod ~ena. Od naaih pet otkrivenih bolesnika etiri su muakarca. Na elferogramu se monoklonski IgD uo ava kao monoklonska vrpca u podru ju gama- ili beta-globulina s popratnom teakom proteinurijom Bence-Jones (2). U opisanom slu aju, kao i u ostala etiri slu aja otkrivenih bolesnika u naaem laboratoriju, uz monoklonski IgD tipa lambda dokazani su BJP tipa lambda u serumu i mokrai, ato se i uklapa u rezultate objavljenih studija koje nalaze da 92% bolesnika razviju BJP tipa lambda (3). Slika hipogamaglobulinemije bila je naena kod svih pet bolesnika. Vrijednosti imunoglobulina G, A i M u naaeg bolesnika jasno pokazuju supresiju njihove sinteze. U opisanog bolesnika nisu naena osteoliti ka ~ariata, oateenje bubrega kao ni organomegalija. S obzirom da je u navedenog bolesnika postignut dobar odgovor na kemoterapiju, odlu eno je da se bolesnik podvrgne lije enju autolognom transplantacijom krvotvornih stanica, ato je i u injeno. Rezultati osnovne laboratorijske obrade, kao i sternalne punkcije nakon navedene transplantacije, pokazali su da je postignuta potpuna remisija bolesti. ABSTRACT - Objective: to present laboratory diagnosis of multiple myeloma by immunoglobulin D (IgD MM). Methods: serum protein electrophoresis on acetate cellulose; immunonephelometric determination of immunoglobulins G, A and M; immunoelectrophoresis for serum monoclonal immunoglobulin D (IgD) determination; immunoelectrophoresis for demonstration of Bence-Jones protein (BJP) in serum and immunofixation to demonstrate urinary Bence-Jones protein. Results: Monoclonal IgD of lambda type and lambda-type BJP were demonstrated in in the patient's serum and urine. Conclusions: (IgD MM) is a rare disease and a cause of only 2% of newly diagnosed cases of multiple myeloma, usually with aggressive course and poor prognosis. Laboratory diagnosis of monoclonal immunoglobulin D should be performed according to Diagnostic algorithm for definition of monoclonal protein elaborated by the Subcommittee for Laboratory Hematology and Morphology, National Group for Treatment of Hematologic Diseases. Key words: IgD multiple myeloma; BJP lambda type; immunofixation LITERATURA: 1. Rowe DS, Fahey JL. A new class of human immunoglobulins I. A unique myeloma protein. J Exp Med 1965;121:171-199. 2. Tan KL, Suri R, Lim L, Ong SK. Immunoglobulin D multiple myeloma in our hospital A rare occurrence. Singapore Med J 2000;41(10):500-503. Lin TL, Shih LY, Dunn P, Wang P, Wu JH, Kao MC. Immunoglobulin D multiple myeloma. Chang Gung Med J 2000;23:451-7. The University of Iowa, Department of Pathology, Laboratory Services Handbook: Immunoglobulin D (http://www.medicine.uiowa.edu/Path_Handbook/handbook/test1091.html) The Community College of Baltimore County. Course on the Adaptive Immune System (http://www.cat.cc.md.us/courses/bio141/lecguide/unit3/humoral/ antibodies/5classes/5classes.html) Dibbern DA, Jr. Immunoglobulin D deficiency, 2002. (www.emedicine.com/med/topic1160.htm) Martin M. Immunoglobulins - structure and function (http://www.microbio.uab.edu/Dental-Opt/Lectures/Martin.pdf)   OPIS SLIKA FIGURE LEGEND Slika 1. Elektroforeza proteina u serumu i imunoelektroforeza seruma i 100 x uguaene mokrae Figure 1. Serum protein electrophoresis and immunoelectrophoresis of serum and 100 x concentrated urine Slika 2. Elektroforeza proteina u serumu i imunofiksacija 100 x uguaene mokrae Figure 2. Serum protein electrophoresis and immunofixation of 100 x concentrated urine Tablica 1. Rezultati laboratorijske obrade Table 1. Laboratory test results (Tablica je u posebnom  fileu jer sam je morala napisati na stranici u polo~aju  landscape !!!!!!!!) 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