DTIC I., I, I 8 8. N LD Lfl 0. N. IELECTE FEB2 8 89D Gordon R. Dreesman HTLV III VIRUS ISOLATION STUDIES ANNUAL REPORT. October 30, 1987.

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1 N LD Lfl 0. N AD HTLV III VIRUS ISOLATION STUDIES Q DTIC ANNUAL REPORT IELECTE FEB2 8 89D Gordon R. Dreesman October 30, 1987 Supported by U.S. ARMY MEDICAL RESEARCH AND DEVELOPMENT COMMAND Fort Detrick, Frederick, Maryland Contract No. DAMD17-86-C-6280 SOUTHWEST FOUNDATION FOR BIOMEDICAL RESEARCH San Antonio, Texas Approved for public release; distribution unlimited. The findings in this report are not to be construed as an official Department of the Army position unless so designated by other authorized documents. I., I, I 8 8

2 Ia. REPORT SECURITY CLASSIFICATION Unclassifi ed Form Approved REPORT DOCUMENTATION PAGE OMe No 0704.l08 lb. RESTRICTIVE MARKINGS 28. SECURITY CLASSIFICATION AUTHORITY 3. DISTRIBUTION/AVAILABILITY OF REPORT Approved for public release: 2b. DECLASSIFICATIONIDOWNGRADING SCHEDULE distribution unlimited 4. PERFORMING ORGANIZATION REPORT NUMBER(S) S. MONITORING ORGANIZATION REPORT NUMBER(S) DAMD17-86-C a. NAME OF PERFORMING ORGANIZATION 6b. OFFICE SYMBOL 7a. NAME OF MONITORING ORGANIZATION Southwest Foundation for (if applicable) Biomedical Research I 6c. ADDRESS (City. State, and ZIP Code) 7b. ADDRESS (City, State, and ZIP Code) San Antonioi TX Be. NAME OF FUNDINGJSPONSORING' 8b. OFFICE SYMBOL 9. PROCUREMENT INSTRUMENT IDENTIFICATION NUMBER ORGANIZATION U.S. Army Medical (if applicable) Research & Development Command DAMD17-86-C-6280 Sc ADDRESS (City, State. and ZIP Code) 10. SOURCE OF FUNDING NUMBERS PROGRAM PROJECT I TASK UWORK NIT Fort Detrick ELEMENT NO. NO. M NO. ACCESSION NO. Frederick, MD M2-9I AB TITLE (Include S*curty Classification) HTLV III Virus Isolation Studies 12. PERSONAL AUTHOR(S) Dreesman, Gordon R. 13a. TYPE OF REPORT 13b. TIME COVERED 14. DATE OF REPORT (Year, Month, Oay) 15. PAGE COUNT 8 6 Annual FROM9/29/ TO 9/28/87 87/10/ SUPPLEMENTARY NOTATION 17. COSATI CODES 18. SUBJECT TERMS (Continue on reverse if necessary and identify by block number) FIELD GROUP SUB-GROUP b U. j RAI, Human immunodeficiency virus ' ABSTRACT (Continue on reverse if necessary and identify by block number) - Virus isolation studies have been performed on 329 blood specimens obtained from Air Force personnel positive for antibody for HIV. Peripheral blood lymphocytes (PBL) from each individual were co-cultured with normal human PHA stimulated PBL. Cultures yielding positive viral reverse transcriptase (RT) activity were scored as positive for HIV. The overall isolation rate for this 12 month study has been 52% (172/329). Patients classified as WRI and WRII yielded a positive virus isolation rate of 51% whereas the collective group with WR classifications of III-VI were 66% positive. Only one positive virus isolation was observed in testing of 40 spinal fluids. Correlation of positive virus isolation with clinical and immunologic parameters is currently being analyzed. 20. D$STRIUTIONIAVAIIASIIITY OF ABSTRACT 21. ABSTRACT SECURITY CLASSIFICATION D]UNCASSIFIfOIUNUMITED Ed SAME AS RPT. 03 OTIC USERS UNCLASSIFIED IA. NAME Of RISPONSiL INDVIDUAL 22b Lf 9 NE _3ZMd Area Code) 22SGMOFFICE SYMBOL MARY FRANCES BOSTIAN i"nrd-rmi- sl S DD Form JUN N Previous edtjons are obsolete. SECURITY CLASSIFICATION OF THIS PAGE

3 Foreword: For the protection of human subjects the investigator(s) have adhered to policies of applicable Federal Law 45CFR46.

4 TABLE OF CONTENTS Page Foreword... 1 Progress Report... 3 Table Table Publications Distribution List

5 Progress Report During the past year we have cultured peripheral blood lymphocytes (PBL) from Air Force personnel who have confirmed positive antibody for HIV. We have also tested a limited number of cerebrospinal fluids for presence of infectious virus. The PBL were processed by Ficoll-Hypaque centrifugation and co-cultured with normal PHA stimulated human lymphocytes for a 3 week period. Media from these cultures were collected at 2 to 3 day intervals starting on day 5 and tested for the presence of viral reverse transcriptase (RT) activity. All cultures which had an increase of threefold greater than control cultures for RT activity were scored positive. Those yielding two- to threefold greater RT activity than uninfected controls were rated at ± and the supernatants were recultured for virus activity. We have completed the testing of 329 blood specimens (Table 1). The overall isolation rate to date has been 172/329 positive (52%) for the 12 month period the contract has been in effect. Table 2 lists isolation results in relation to the Walter Reed status of the tested individuals. Class 1 and 2 specimens still predominate and the isolation remains around 50% in both groups. The numbers of Class 3 through 6 are beginning to increase, but still remain relatively low. Although the significance of the findings in these groups is questionable due to the small numbers, the overall isolation rate for these later four groups is greater than that found in the earlier stages (66%). The new laboratory that is committed to the performance of this contract is now completed and has been occupied. The computer terminal, which was installed during the last quarter, is now in use to transfer specimen testing results to patient files at Wilford Hall USAF Medical Center. It has also allowed us to check the Walter Reed classification of patients not previously reported. During the next year we will analyze the individuals with positive cultures to see if there are any clinical or immunologic parameters which might serve as an indicator of virus expression in asymptomatic patients. The installation of the computer terminal will greatly facilitate this analysis. Only one of 40 spinal fluids has yielded a positive virus culture in a limited study this past year. We are currently testing freshly collected cerebrospinal fluids to determine the validity of the negative results obtained from earlier specimens which have been stored frozen for some time prior to testing. Previously, the spinal fluids were received frozen. In our current test protocol we are attempting to isolate virus by two methods: 1) we are co-cultivating cells isolated from fresh spinal fluid with PHA stimulated normal human PBL and 2) 1 ml of clarified fresh fluid will be used to resuspend 0.5 x 106 CEM A3.01 target cells containing a final concentration of 10 ng/ml polybrene. 3

6 TABLE I HEY ISOLATION FROM LYMPHOCYTES... RT RESULTS DATE OF DAY 5 DAY 7 DAY 10 DAY 12 DAY 15 DAY 18 DAY 21 POS/ WR PATIENT# COLLECTION CPM CPM CP4 CPM CPN CPM CPN NEG STATUS... WH2/ NT POS UH3/ POS WH4/ NT 7863 NT NT POS WH6/ MEG WH23/ * POS WH24/ NT NEG WH26/ MEG W,'H28/ NT MEG WH30/ MEG WH32/ POS W, H33/ NT NT MEG WH43/ NEG BANC34/ POS BAMC35/ MEG WH/B POS UH/N POS 2 01/15/ POS 3A 2/2 06/6/ MEG 3/2 06/06/86? MEG 15 03/26/ POS /21/ POS 5B 39 08/13/ POS 2AITP 42 11/26/ MEG 1A 51 12/18/ MEG 2A 54 07/30/ POS 3A 55 12/04/ MEG 3A 66 10/23/ NEG(NEG) 2A 66 11/13/ MEG 2A 69 02/19/ POS IA 74 02/19/ POS 3A 75 05/14/ MEG IA 76 08/20/ MEG 5A 80 02/19/ MEG IA 81 08/27/ /-POS /28/ POS IA 90 07/16/ POS 2A 95 05/14/ POS 1A 96 08/20/ MEG IA 99 05/21/ POS 1A /21/ POS 2A /16/ POS 5A /09/ MEG 1A 4

7 113 08/27/ MEG 3A /30/ POS 2A /13/ POS 3A /11/ POS 3A /30/ POS 1A /20/ MEG 2A /30/ POS 2A /30/ MEG 1A /20/ MEG 1A /13/ MEG 2A /23/ NEG(MEG) 2A /23/ NEG(NEG) 2A /23/ NEG(POS) 1A /23/ NEG(POS) 1A /30/ NEG(NEG) /30/ NEG(NEG) 1A /30/ NEG(NEG) 2A /30/ NEG(POS) 2A /13/ MEG 2A /30/ NEG 5B /30/ NEG(POS) IA /30/ NEG(POS) 3A /13/ POS 3A /30/ NEG(POS) 2A /13/ MEG 1A /30/ NEG(POS) 1A /30/ NEG(POS) 2A /30/ MEG 2A /06/ MEG 1A / POS 1A /06/ POS 1A /06/ POS 1A /06/ POS 5A /06/ MEG 2A /06/ POS 1A /06/ POS 1A /06/ POS 1A /06/ MEG IA /13' MEG 1A /13/ MEG 3A /13/ MEG 1A /13/ MEG 1A /20/ MEG IA /20/ MEG 1A /20/ MEG 1A /20/ POS 2A /20/ MEG 1A /20/ MEG 1A /20/ MEG IA /26/ POS 1A /20/ MEG 1A /20/ MEG IA 5I

8 217 11/25/ NEG 1A /26/ NEG 3A /26/ NEG 1A /26/ NEG 2A /26/ NEG 1A /26/ POS 2A /26/ POS 1A /26/ NEG IA /26/ POS 4A /04/ POS 1A /04/ POS 2A /04/ POS 1A /04/ NEG IA /04/ POS 5A /04/ NEG IA /04/ NEG IA /11/ /-POS 1A /11/ POS IA /11/ NEG 1A /11/ NEG 1A /11/ POS 1A /11/ /-POS /11/ NEG 1A /11/ NEG 1A /18/ POS 1A /18/ POS IA /18/ POS 1A /18/ POS 1A /18/ POS IA /18/ MEG 1A /18/ /-POS 1A /18/ NEG 2A /18/ POS /23/ NEG 1A /23/ NEG IA 264 BABY 12/23/ NEG /23/ POS 1A /23/ POS 2A /23/ NEG IA /23/ MEG IA /23/ NEG IA /23/ POS IA /23/ /-POS IA /23/ MEG 1A /30/ NEG IA /30/ NEG IA /30/ NEG 1A /08/ /-POS 1A /08/ /-POS 1A /08/ /-POS IA /08/ POS IA /08/ POS 1A 6

9 281 01/08/ MEG IA /18/ MEG 1A /18/ MEG 1A /18/ MEG 2A /15/ NEG IA /15/ /-POS 2A /15/ POS 1A /15/ /-POS 1A /22/ POS 1A /22/ POS 1A /22/ POS 1A /22/ POS 1A /22/ MEG 1A /29/ POS 1A /29/ POS 1A /29/ POS 1A /05/ POS 1A /05/ POS 1A /05/ POS 5A /05/ POS 1A /05/ POS 1A /05/ POS 1A /05/ MEG 1A /05/ POS 3A /12/ POS IA /12/ POS IA /12/ POS 1A /12/ MEG 1A /12/ , MEG IA /12/ POS 2A /12/ MEG 1A /19/ MEG 1A /19/ POS 1A /19/ POS 1A /19/ POS 1A 32-02/19/ POS 1A /19/ MEG 1A /26/ MEG IA /19/ POS 1A /19/ MEG 1A /26/ POS 1A /19/ POS 1A /26/ POS 2A /26/ POS 1A /26/ POS 1A /26/ PO$ /19/ POS 1A /05/ MEG 2A /05/ POS 1A /05/ /-POS 1A /05/ MEG 1A /05/ MEG 1A 7

10 343 03/05/ POS IA /19/ MEG 2A /12/ POS 1A /12/ POS 1A /12/ MEG 2A /12/ POS IA /12/ MEG 1A /12/ Pos 2A /12/ POS 1A /12/ MEG 2A /12/ POS 2A /12/ POS 1A /19/ POS 2A /26/ POS /19/ POS 2A /19/ POS 1A /26/ POS /19/ /-POS 2A /26/ MEG 2A /26/ POS 1A /26/ MEG 1A /26/ POS 1A /26/ POS IA /26/ POS 2A /26/ MEG 2A /26/ MEG 1A /26/ POS IA /26/ MEG 1A /09/ POS 1A /09/ POS 1A /09/ POS 2A /09/ POS 2A /09/ POS 1A /09/ MEG IA /09/ /-POS 1A /16/ MEG 1A /16/ POS 1A /16/ i 1.8 MEG 1A /16/ POS 1A /16/ MEG IA /16/ MEG 1A /23/ POS 1A /23/ POS 1A /23/ MEG IA /23/ /-POS IA /30/ POS IA /30/ POS 1A /30/ /-POS 1A /07/ MEG 1A /30/ POS IA /14/ POS 2A /07/ MEG IA 8

11 412 05/07/ POS 1A /07/ NEG 2A /07/ NEG 1A /07/ NEG 1A /07/ NEG 1A /14/ POS /07/ MEG IA /14/ NEG 1A /14/ POS 1A /14/ POS 1A /21/ POS 1A /21/ NEG 1A /21/ POS 1A /28/ POS 1A /28/ POS 1A /04/ NEG 2A /28/ NEG IA /28/ POS 1A /28/ U POS 1A /04/ u NEG 2A /04/ POS 1A /04/ NEG 1A /04/ POS 2A /11/ POS /11/ NEG 2A /18/ POS /11/ POS IA /18/ POS 1A /18/ POS 1A /18/ POS 1A /18/ POS 3A /18/ NEG 1A /18/ NEG 1A /18/ NEG IA /25/ NEG IA /25/ POS IA /25/ POS 1A /25/ POS 1A /09/ NEG 5A /02/ POS 4A /02/ POS 1A / ND NEG 1A /09/ /-POS 1A /02/ POS 3A /02/ NEG /09/ POS IA /09/ POS IA /16/ POS 1A /16/ POS 1A /16/ POS 3A /16/ POS IA /16/ POS IA 9

12 547 07/23/ /-POS 1A /23/ NEG 1A " NEG 1A /23/ POS 1ACHL /23/ POS 1A RH 07/23/ NEG KW 07/23/ NEG /30/ POS 1A /30/ POS 1A /30/ POS 1A /30/ POS 1A /30/ POS 1A /30/ NEG 1A /06/ ND POS 1A /06/ ND POS IA /06/ ND POS IA /27/ NEG /13/ POS 1A /13/ POS 4AK /27/ NEG 1A /20/ NEG 1A /20/ /-POS 1A /20/ NEG 1A /20/ NEG 1A /20/ NEG 1A /20/ /-POS 1A /20/ MEG 1A * AVG CPN/BACKGROUND CPH; >3.0 = POS CULTURE; >2.0 = /'POS 10

13 TABLE 2 HIV ISOLATION FROM LYMPHOCYTES BY WALTER REED CLASSIFICATION Walter Reed Classification Positive Negative (52%) 103 (48%) 2 24 (44%) 31 (56%) (66%) 11 (33%) Total 157 (52%) 145 (48%) 0i 0! 11!

14 DISTRIBUTION LIST 4 copies Director Walter Reed Army Institute of Research Walter Reed Army Medical Center ATTN: SGRD-UWZ-C Washington, DC copy Commander US Army Medical Research and Development Command ATTN: SGRD-RMI-S Fort Detrick, Frederick, Maryland copies Defense Technical Information Center (DTIC) ATTN: DTIC-DDAC Cameron Station Alexandria, VA copy Dean School of Medicine Uniformed Services University of the Health Sciences 4301 Jones Bridge Road Bethesda, MD copy Commandant Academy of Health Sciences, US Army ATTN: AHS-CDM Fort Sam Houston, TX