Impact of HLA-B*35 alleles on HIV disease outcome in Mexico and Central America Valenzuela-Ponce H 1, Ávila-Ríos S 1, Garrido-Rodríguez D 1, García-Téllez T 1, Soto-Nava M 1, Escamilla-Gómez T 1, García-Morales C 1, Cortés-Álvarez J 1, Hernández R 1, Murakami A 1, Mejía- Villatoro C 2, Pascale J 3, Porras G 4, Quant C 4, Lorenzana I 5, Meza R 5, Palou E 5, Manzanero M 6, Reyes-Terán G 1 for the Mesoamerican HIV Project Group. 1 Instituto Nacional de Enfermedades Respiratorias, Mexico; 2 Hospital Roosevelt, Guatemala; 3 Instituto Conmemorativo Gorgas de la Salud, Panama; 4 Hospital Metropolitano Vivian Pellas, Nicaragua; 5 HIV National Program, Honduras; 6 Ministry of Health, Belize. The authors declared no conflict of interest.

Background It is well established that human leukocyte antigen (HLA) influences the rate of HIV disease progression, the most consistent associations have been reported for HLA-B alleles. HLA-B*35 has been associated with accelerated progression with the most deleterious effects linked to a group of subtypes, HLA-B*35-PX. Allele Epitope anchor preference P2P9 (F-pocket) B*35:02PX B*35:03PX B*35:04PX B*35:01PY B*35:08PY X= leucine, methionine or valine, but not tyrosine B*35-PX B*35-PY Reduced rate of disease progression Rapid rate of disease progression 976 ART-naïve, HIV-1 clade B Mexican individuals. No impact on viral load was observed in associations with the B*35-PX/PY groups. However, we observed differences in HIV disease outcome associated with specific B*35 alleles. Carrington M et al, Science 1999.Gao X et al, N Engl J Med Valenzuela-Ponce H et al. AIDS 2014

Cohort characteristics Table 1. Clinical and demographic characteristics CountryMexicoCentral AmericaAll N Age (median, [IQR])30 [24 – 38]33 [26 – 42]32 [ ] Female [N (%)]309 (21.4%)504 (36.0%)813 (28.6%) Log Plasma Viral Load (RNA copies/mL, median [IQR]) 4.68 [4.09 – 5.23]4.60 [3.89 – 5.16]4.64 [4.00 – 5.21] CD4+ T cell counts (cells/μL, median [IQR]) 337 [135 – 555]315 [124 – 504]337 [131 – 541] Mexico (n=1474) 21% ♀, 4.68, 337 Country (n) % ♀ (females), median pVL, median CD4 count Belize (n=102) 49% ♀, 4.3, 470 Honduras (n=340) 42% ♀, 4.58, 300 Nicaragua (n=243) 20% ♀, 4.8, 316 Panama (n=297) 25% ♀, 4.34, 373 Guatemala (n=418) 44% ♀, 4.71, % of the individuals are B*35+

High diversity of relative unstudied B*35 alleles: 27 different alleles, 10 with N>10 PY PX

Diverse ranking of HLA-B*35 alleles according to the median pVL was observed B*35:12 is a novel Amerindian HLA allele associated with higher pVL. PY- PX-

We observed B*35 additive detrimental effect and no homozygous disadvantage for specific B*35 alleles Double postive individuals for other B*35 alleles were excluded from the analysis

No significant impact on median pVL was observed between HLA-B*35-PX/PY groups Allele Residue in HLA peptide binding groove Epitope anchor preference P2P9 B*35:02NYPX B*35:03-FPX B*35:04--PX B*35:12--PX 3 other B*35-F/YPX B*35:01DSPY B*35:08--PY B*35:14--PY B*35:16--PY B*35:17--PY B*35:20--PY B*35:43--PY 13 other B*35--PY x= leucine, methionine or valine, but not tyrosine B*35-PX Putative B*35-PX B*35-PY Putative B*35-PY

The magnitude of B*35 alleles effect in each region/country was not frequency-dependent

Summary B*35:01 (PY), B*35:02 (PX) and B*35:12 (which is not considered disease-susceptible allele), were associated with higher pVL (p<0.05, q<0.1). We observed B*35 additive detrimental effect and no homozygous disadvantage for specific B*35 alleles. Our data indicate that the classification of HLA-B*35 sybtypes into PX and PY groups does not satisfactorily explain differences in median pVL. These results challenge the B*35-PX/PY hyphothesis, indicanting that PY alleles can be disease-susceptible and also that differences exist in disease associations within the PX/PY grouping. Also the previous observation that the negative effect of the B*35 allelic group is due to PX alleles is not supported by these data. We conclude that the detrimental effect of B*35 is unlikely to be related exclusively to PX/PY grouping.

Acknowledgments Funding : Mexican Government (Comisión de Equidad y Género de la H. Cámara de Diputados). National Council of Science and Technology (CONACYT). Gustavo Reyes Terán Santiago Ávila Ríos CIENI Center for Research in Infectious Diseases, Mexico City HIV-infected blood donors Daniela Garrido Selma Alva Maribel Soto Tania Escamilla Claudia García Thalía García Jonathan Cortés Silvia del Arenal CIENI students Collaborators/HLA typing Ramón Hernández Edna Rodríguez Mario Preciado Carolina Demeneghi Raymundo González Israel Molina Clinical Lab Zeidy Arenas Sandra Zamora Berenice Cancino Administrative staff Pablo César Estebán Paz Informatics staff CIENI medical team Akio Murakami María Gómez-Palacio Karla Romero  Dra. Indiana Torres (Puebla)  Dr. César Carrasco (Oaxaca)  Dr. Jaime Andrade (Jalisco)  Dra. Sara Aguirre (Jalisco)  Dra. Lucero González (Jalisco)  Dra. Beatriz Ramírez (Edo. México)  Dr. David de los Santos (Guerrero)  Dr. Juan Carlos Martínez (Guerrero)  Dra. Adakatia Armenta S. (Guerrero)  Dr. Samuel Wong González (Sonora)  Dra. Angélica Zimbrón (Baja California)  Dr. Samuel Navarro (Baja California)  Dra. Lucrecia Ceja (Nuevo León)  Dra. Ma. Catalina Juárez U. (Nuevo León)  Dr. Gastón Coronel (Veracruz)  Dr. Yolanda Chévez (Morelos)  Dra. Adriana García (Morelos)  Biol. Anselmo Hernández C. (Morelos)  Dra. Margarita Aguilar Ruiz (Chiapas) Epidemiology Project Group Physician from Mexican states: Collaborators from Central America Dr. Mexía Villatoro C (Guatemala) Dr. Pascale J (Panama) Dr. Porras G (Nicaragua) Dr. Quant C Nicaragua) Dr. Lorenzana I (Honduras) Dr. Meza R (Honduras) Palou E (Honduras) Manzanero M (Belize)