Guadalupe Ayora-Talavera

Mutations in H5N1 Influenza Virus Hemagglutinin that Confer Binding to Human Tracheal Airway Epithelium

The emergence in 2009 of a swine-origin H1N1 influenza virus as the first pandemic of the 21st Century is a timely reminder of the international public health impact of influenza viruses, even those associated with mild disease. The widespread distribution of highly pathogenic H5N1 influenza virus in the avian population has spawned concern that it may give rise to a human influenza pandemic. The mortality rate associated with occasional human infection by H5N1 virus approximates 60%, suggesting that an H5N1 pandemic would be devastating to global health and economy. To date, the H5N1 virus has not acquired the propensity to transmit efficiently between humans. The reasons behind this are unclear, especially given the high mutation rate associated with influenza virus replication. Here we used a panel of recombinant H5 hemagglutinin (HA) variants to demonstrate the potential for H5 HA to bind human airway epithelium, the predominant target tissue for influenza virus infection and spread. While parental H5 HA exhibited limited binding to human tracheal epithelium, introduction of selected mutations converted the binding profile to that of a current human influenza strain HA. Strikingly, these amino-acid changes required multiple simultaneous mutations in the genomes of naturally occurring H5 isolates. Moreover, H5 HAs bearing intermediate sequences failed to bind airway tissues and likely represent mutations that are an evolutionary “dead end.” We conclude that, although genetic changes that adapt H5 to human airways can be demonstrated, they may not readily arise during natural virus replication. This genetic barrier limits the likelihood that current H5 viruses will originate a human pandemic.

Receptor Binding Profiles of Avian Influenza Virus Hemagglutinin Subtypes on Human Cells as a Predictor of Pandemic Potential

ABSTRACT The host adaptation of influenza virus is partly dependent on the sialic acid (SA) isoform bound by the viral hemagglutinin (HA). Avian influenza viruses preferentially bind the α-2,3 SA and human influenza viruses the α-2,6 isoform. Each isoform is predominantly associated with different surface epithelial cell types of the human upper airway. Using recombinant HAs and human tracheal airway epithelial cells in vitro and ex vivo, we show that many avian HA subtypes do not adhere to this canonical view of SA specificity. The propensity of avian viruses to adapt to human receptors may thus be more widespread than previously supposed.

Human papillomavirus infection and spontaneous abortion: a case–control study performed in Mexico

OBJECTIVE To investigate if HPV cervical infection is associated with spontaneous abortion in a Mexican population. STUDY DESIGN Case control study including 281 women from two Social Security Hospitals in Merida, Mexico. Cases were women with spontaneous abortion attending for curettage, and controls were pregnant women at term who attended for delivery. HPV molecular detection and typing of HPV 16, 18, 58 and 6/11 was performed on cervical samples, and TORCH serology IgM tests (against T. gondii, CMV, HSV) were performed on cases. Data were analyzed using Chi square, odds ratio and linear regression tests. RESULTS HPV global prevalence was 19.8% (24.4% in cases and 15.2% in controls). HPV types 16 and 58 were the most frequently detected in both groups. Multiple HPV types concurrent infection were found in 31.4% of typified samples. Amongst cases 27.3% of HPV positive women reported at least one previous pregnancy loss; compared to 17.43% amongst HPV negative women. Nevertheless, HPV was not significantly associated with spontaneous or to repetitive abortion. Cases were 60.2% positive to any TORCH agent, although it was not significantly associated to referred miscarriage history. Spontaneous abortion was associated to a previous pregnancy loss and to womens age older than 35 years old. HPV infection was significantly associated to alcohol intake before pregnancy and to multiple sexual partners. CONCLUSION HPV cervical infection was not associated with spontaneous abortion. HPV in spontaneous abortion and other adverse pregnancy outcomes merits further study.

Human papillomavirus 13 in a Mexican Mayan community with multifocal epithelial hyperplasia: could saliva be involved in household transmission?

Multifocal epithelial hyperplasia (MEH) is a disease of the oral mucosa. Human papillomaviruses 13 and 32 have been detected in these lesions. We describe the epidemiology and clinical characteristics of patients with MEH in a rural community in the Mayan area of Mexico with 53 cases and 54 controls. Clinical and epidemiological data were collected through a direct interview. Oral cell samples were collected with a cytobrush. Subjects collected their own saliva sample in a sterile bottle. All samples were tested for HPV 13 and 32 by polymerase chain reaction using specific primers. Of the 53 patients and 54 healthy subjects, 56% were < 12 years old, 25% were males and 75% females. Evolution of the lesions was between two months and 17 years. The lesions affected lips, jugal mucosa, and tongue, 96% had multiple lesions. From 53 patients, fifty samples of oral cells and 31 samples of saliva were analyzed. HPV 13 was detected in 100% oral cell and 100% saliva samples studied. 16 healthy subjects were HVP 13 positive. A highly significant association of HPV 13 infection and MEH was found, as determined by chi square test (p = 0.00) Household transmission of HPV 13 may happen through saliva and the shared use of contaminated objects.

Natural Vertical Transmission of Dengue-1 Virus in Aedes aegypti Populations in Acapulco, Mexico

Abstract We carried out dengue virus surveillance in Aedes aegypti populations from 47 neighborhoods of Acapulco during the rainy season of 2011 following a standard national protocol and as an improvement of the entomological surveillance of the Mexican Dengue Control Program. A total of 4,146 Ae. aegypti adults collected indoors and/or emerged from eggs, larvae, or pupae from households with dengue reports (probable or confirmed cases), were grouped into pools and processed using a standardized serotype-specific 4-plex real-time reverse transcriptase–polymerase chain reaction assay. Overall, only 2 (0.9%) of 226 pools of Ae. aegypti adults (1 pool of adults emerged from field-collected larvae, and another of indoor-collected adults) were positive for dengue virus 1 (DENV-1). This is appears to be the 1st report of evidence on the vertical and transovarial transmission of DENV-1 in field-caught Ae. aegypti in Mexico.

Amino acid substitution D222N from fatal influenza infection affects receptor-binding properties of the influenza A(H1N1)pdm09 virus

We have analyzed the receptor binding profile of A(H1N1)pdm09 recombinant influenza viruses containing the amino acid substitution D222N which has been associated with a fatal case of infection. This mutation was investigated in conjunction with a secondary mutation, S185N. Using human tracheobronchial epithelial cells (HTBE), we found that single mutation D222N affects the binding and replication of the virus during initial stages of infection, with limited but preferred tropism to non-ciliated cells expressing α2,6-SA. However, in conjunction with the S185N change, the (D222N, S185N) virus shows a remarkable increase in binding and replication efficiency, with tropism for both ciliated and non-ciliated cells. Glycan microarray analysis demonstrated correlation between the binding profile and the cell tropism observed in the HTBE cells. These findings suggest that viruses with D222N required compensatory mutations such as S185N to maintain viral fitness, and in combination, affect the pathogenicity of the virus and the clinical outcome.

Epidemiology of oral HPV in the oral mucosa in women without signs of oral disease from Yucatan, Mexico

High-risk human papillomaviruses (HR-HPV) are considered necessary for the development of cervical cancer. Furthermore, there is no doubt that some types of oral squamous cell carcinoma are associated with HR-HPV. The epidemiology of oral HPV infections in healthy subjects remains unclear due to a lack of knowledge. The objective of this study was to investigate the epidemiology of human papillomavirus infections of the oral mucosa without pathology. A cross-sectional study was performed; samples from 390 women seeking prenatal care, Pap smears, family planning or gynecological diseases were studied. Oral cells were collected by direct swab sampling. Information regarding sociodemographic status, sexual behavior, infectious diseases, contraceptive history and tobacco and alcohol consumption were obtained through direct interviews. HPV and genotypes were detected by type-specific polymerase chain reaction. Our results revealed that 14% of the women studied had an oral HPV infection. Women ≤ 20 years of age had the highest HPV prevalence (24.5%). In total, seven genotypes were identified, including the high-risk genotypes 16, 18, 58 and 59 and the low-risk genotypes 6, 81 and 13, the latter of which is a type exclusive to oral mucosa. Sexual behavior was not associated with the presence of genital HPV types in the oral mucosa. Genital HPV types were present in the oral mucosa of women without associated clinical manifestations; however, sexual behavior was not associated with infection, and therefore others routes of transmission should be explored.

Arbovirus Surveillance and First Report of Chikungunya Virus in Wild Populations of Aedes aegypti from Guerrero, Mexico

Abstract We carried out dengue (DENV) and chikungunya virus (CHIKV) surveillance in wild populations of Aedes aegypti from Guerrero, Mexico, from 2012 to 2014 following a standard national protocol of the Mexican Dengue Control Program. A total of 284 pools (15–30 specimens/pool) of female mosquitoes were tested with real-time reverse transcriptase–polymerase chain reaction to detect DENV and CHIKV. We report for the 1st time the detection of CHIKV from field-collected mosquitoes at Acapulco and Juchitán in 2014. Results from DENV are also reported.

Hemagglutinin variants of influenza A(H1N1)pdm09 virus with reduced affinity for sialic acid receptors

In influenza A H1 virus, amino acids at position 190 and 225 of HA affect replication and transmission. In this study, we show that the mutation D190Y in the HA of influenza AH1N1pdm09 virus reduces the affinity of the virus for sialic acid receptors expressed at the surface of red blood cells from different species without affecting virus replication in MDCK cells.

The Risk of Aedes aegypti Breeding and Premises Condition in South Mexico

Abstract A recent innovation instrumented for the Dengue Prevention and Control program in Mexico is the use of the premises condition index (PCI) as an indicator of risk for the vector Aedes aegypti infestation in dengue-endemic localities of Mexico. This paper addresses whether further improvements for the dengue control program could be made if the prevalence and productivity of Ae. aegypti populations could be reliably predicted using PCI at the household level, as well as medium-sized neighborhoods. We evaluated the use of PCI to predict the infestation with Aedes aegypti (breeding sites and immature productivity) in Merida, Mexico. The study consisted of a cross-sectional survey based on a cluster-randomized sampling design. We analyzed the statistical association between Aedes infestation and PCI, the extent to which the 3 components of PCI (house maintenance, and tidiness and shading of the patio) contributed to the association between PCI and infestation and whether infestation in a given premises was also affected by the PCI of the surrounding ones. Premises with the lowest PCI had significantly lower Aedes infestation and productivity; and as PCI scores increased infestation levels also tended to increase. Household PCI was significantly associated with Ae. aegypti breeding, largely due to the effect of patio untidiness and patio shade. The mean PCI within the surroundings premises also had a significant and independent explanatory power to predict the risk for infestation, in addition to individual PCI. This is the 1st study in México showing evidence that premises condition as measured by the PCI is related to Ae. aegypti breeding sites and immature productivity. Results suggest that PCI could be used to streamline surveys to inform control efforts at least where Ae. aegypti breeds outdoors, as in Merida. The effect of individual premises, neighborhood condition, and the risk of Aedes infestation imply that the risk for dengue vector infestation can only be minimized by the mass effect at the community level.