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Pilot Study of the Effects of Thymus Protein on Elevated Epstein-Barr Virus Titers by Neil H. Riordan, PA-C, BS1;James A. Jackson, PhD, MT (ASCP),CLS, BCLD2;and Hugh D. Riordan, MD3 Project RECNAC Bio-Communications Research Institute 3 100 N. Hillside Ave. Wichita, Kansas 67219 Financud Support Pcnfied pmteln A from calf thymus cell culture (BloPm Thym~cProte~nATM).was a plfl fmm LOngewty Sc~enca,New York, New York lymphoproliferative d i ~ o r d e r s and ,~ neurological disorder^.^ In addition to primoinfection and reactivation, a chronic-active EBV infection has been described." EBV infection occurs more frequently in immunocompromised hosts?1° and has been reported to be immunopathologic in presumably immunocompetent h ~ s t s . ' A ~ cytOtoxic .~~ T-lymphocyte (CTL) response i s responsible for resolution of EBV infections.l3 Purified protein A from calf thymus cell culture (BioProThymic Protein ATM) is a patented, over-the-counter supplement which contains a polypeptide that has shown the ability Background to stimulate mature T lymphocytes and enhance mammalian immune response Epstein-Barr virus (EBV) infection 1 is extremely common, approaching to infectious agents a n d to malignancies.14 This polypeptide was 100% by the age of 10 in some parts of the world. EBV is thought to be spread first studied a t UCLA in 1979. Since that via the respiratory route, and is the time, a bovine thymus cell line which produces t h e protein h a s been causative a g e n t of infectious rnononucleosi~.~ established. The protein is chromatographically purified from a After acute infection, EBV has been I shown to establish a latency state within medium used to culture the cell line. It is now commercially available in a base B lymphocytes, salivary gland of maltodextrin, a n d is t a k e n epithelium, a n d oropharyngeal epithelial cells.2 Like other herpes sublingually (4 mcg/dose). viruses, EBV can reactivate. Quantitation of IgG antibodies to Primoinfection and reactivation of EBV EBV early antigen (EBV EA) is have been associated with several commonly used to detect recent or chronic-active infection. Elevated serum disease states, including chronic fatigue syndrome (CFS), Burkitt's l y m p h ~ m a , ~ viral capsid (EBV VCA) antibodies indicate previous infection and can I nasopharyngeal carcinoma,' aplastic anemia,' Hodgkin's disease, several remain elevated for life. 1 Abstract S i x p a t i e n t s with chronically e l e v a t e d Epstein-Barr virus early a n t i g e n a n t i b o d y t i t e r s w.ere t r e a t e d f o r 60 d a y s with purified P r o t e i n A f r o m calf t h y m u s cell c u l t u r e (BPTPA). T h e r e w a s a statistically significant reduction of antibody titers after treatment. T h e t h y m i c protein was well tolerated a n d i n c r e a s e d energy was reported b y m o s t of the participants. These resulta suggest t h a t BPTPA may be useful in the treatment of patients with persistently elevated EpsteinB a r r virus e a r l y antigen antibody titers. 1 ' 1 Objective The objective of this study was to determine what effect BPTPA, a T lymphocyte stimulant, had on chronically elevated EBV EA titers. Methode Seven patients meeting the criterion of having persistent (>6 months) EA titers of greater than or equal to 1:80 (Negative <1:20) were recruited to participate in the study for 60 days. Participation was entirely voluntary, and informed consent was received from all participants. The study protocol was approved by our Institutional Review Board and was carried out with the ethical standards s e t forth in the Helsinki Declaration of 1975. Participants were provided with 4 mcg. packets of BioPro Protein& which were taken sublingually,three times per day, for a period of 60 days. At the end of the study period, EBV EA and EBV VCA levels were re-tested. Participants were also requested to complete a symptom questionnaire before a n d after t h e study. The questionnaire contained 86 current symptom questions which were ranked by the participants on a scale of 1-7 (lkno symptom, 7=severe symptom). The pre- and post-treatment scores were compared using the student T-test. -Determinations of EBV EA ( 1 s ) and EBV VCA (IgG) were performed by Laboratory Corporation of America, Wichita, Kansas. EBV EA (Negative: 1 78 TOWNSEND LElTEA for DOCTORS L PATIENTS - F E ~ A U A R Y ~ M A R C1998 H a -. 13 'C] 5 - 2 p I <1:20) was quantified using indirect fluorescent antibody, and EBV VCA (Negative: <20 arbitrary units) was determined using enzyme immunoassay. The pre- and posttreatment scores for both assays were compared using the student T-test. Reeults Of the seven patients enrolled, six completed the study. One patient discontinued use after 7 days after experiencing flu-like symptoms which resolved within three days of discontinuation. No adverse side effects were reported by t h e other six participants. Four participants completed both before and after symptom questionnaires. Compliance was good, with greater than 90% of dosea taken. Table 1 shows the ages, sexes, and diagnoses of the six participants. T h e results of t h e study a r e summarized in Table 2. After treatment the EBV EA levels of four of the six participants were reduced. The reduction waa s&nXcaat(pe.03). There was no reduction of the EBV VCA levels after treatment. The overall trend of the symptom profiles was toward a reduction in symptoms. Of the four before and after profiles completed, two showed 1 significant (p=.04) reduction i n symptoms after treatment (Table 3). 1 - In poet-study i n t e ~ e w sall , of the 5 participants with a diagnosis of CFS reported increased energy and a decrease in houm slept after the 60 day treatment. Also of note was that elevated liver enzymes (ALT, AST, and GGT), and cholesterol in patient number 3, who had a concomitant diagnosis of hepatitis C, and leukopenia in patient number 4, were all normalized by the end of the study. Dhxssion Although this was a non-blinded pilot study, the fact that a significant reduction of EBV EX titers occurred in a group of patiente who had persistently elevated titers suggests that BPTPA may play a role in modulation of chronicactive EBV infection. References 1. 2. 3. 4. 5. 6. 7. 8. 9. lo. Conclusion The role of BPTPA in the management of fatigue and chronicactive EBV infection should be studied further. 11. 12. Correspondence: Neil H. Riordan, PA-C , Manager, Project RECNAC Bi&mmunications Research Institute 3100 N. Hillside Ave. Wichita, Kansas 67219 USA 316-682-3100 Fax 316-682-5054 13. 14. : .. . - 4 1:320 1:SO 1:320 5 . 6 t-test pC03 significant 1. Manager, Project RECNAC. BioGommunications Reaearch Institute. 3100 N. Hillside Ave.. Wichita. Kansas 67219 2. Professor, Wichita State University, Department of Medical Technology, Box 43, Wichita, Kansas, 67260-0043. 3. Director, Project RECNAC, Bio-Communications Research Institute, 3100 N. Hillside Ave., Wichita. Kansas 67219 1:80 1:80 1:80 I70 170 1 70 - . Evans AS. Epstein-Barr and infectious mononucleoaia:N Engl J Med. 1972;286:836-37 Simon MW M+festationa of relapsing EpskinBarr virua illness. J Ky Med Assoc 1997 . Jun:95(6):240.243. Stein JH. Sfeini Internal Medicine. 4th ed. St. Louia. Mo: Mosby Yearbook;1994:905. Banatvda JE, Beet JM.Waller D K Epstein-Ban virus-specific IgM in infectious mononucleosis. Burkitt's lymphoma and nasopharyngeal carcinoma. Lancet 1972;i:1205-08. Stein J K Steini I n k m l Medicine. 4th ed. S t . Louis. Mo: Moaby Yearbook;1994:874. Lamy ME. Favart AM. Cornu C et al. Epstein-Barr virus infection in 59 orthotopic liver transplant patients. Med Microbial Immunol (Berl). 1990;179(3):137-144. Liaw SB, Shen EY. Alice in Wonderland syndrome as a presenting aymptom of EBV infection.Pediofr N e u r d 1991Nov;7(6):464466. Buchwald D.Sullivan JL,KomnmBAL. Frequency of chrunic active Epatein-Bam virua infection in a general medical practice. JAMA. 1987;257:23032307. Purtilo DT, Paquin LA, Sakamoto K, e t al. Persistent transfusion associated infectious mononucleosis with transient acquired immuaodetiaency. Am J Mcd 1980SE:437-40. &rnalE G. Uj M.Sacs G, Bali I . h a a i I, Kocai J. Seroiogic a i g o~ f Epstein-Barnvirus infections in various patients without infedioua mononucleoeii. ON He& 11996,137(30):1633-1636. S c h d L. N e m - H a d e l i n D. Persistent (duonic active) Epstein-Bam v i r u ~infection and tuthritis in c h i l d h e M o d KindcrheilM. 196!5;133(11):846-847. Hotehin NA,Read R, Smith DG. Crawford DH. Active Epstsi-Barr virus infeetion in post-viral fatigue syndrome. JInfaci. 1989;18(2):143-150. Morgsn SM. WLinson GW.Floettmann E. Blake N, Rickinson AB. A recombinant adenovirus expmsing an Epstei-Barr virus (EBV) target antigen can selectivelyrendivate rarewmponentn of EBV Eytotoric T-lymphocytememory in vitm. J V i 1996;70(4):2394402. Beardsley, TR. Immune-enhancing agent for therapeutic use in immunocompmmised hosts. US. Patent. 1997; Number5.616.554. 138 170 170 p-. 88 no! sip@an!