HIV-infected infants had resistance profiles largely resembling that of their mothers. Since some of the 17 mothers with ART resistance had multiple mutations, 29 total mutations were observed. Maternal–infant concordance by drug class was as follows. Of five maternal NRTI-associated mutations, three pure mutants corresponded with mutant in the infant (in pairs 8 and 24), while two mixtures corresponded with wild type in the infant (pairs 11 and 17); in both mixtures, wild type was the predominant variant. One NNRTI-associated mutation (K103N) and two major PI-associated mutations (L90M and V82I/V) were observed among mothers (pairs 27 and 15). None of these was transmitted. However, 17 of 21 maternal minor PI-associated resistance mutations (81%) were detected in the corresponding infant isolates; of 16 pure mutations in maternal virus, 15 were identical in infant virus, while for five mutant/wild-type mixtures only the quantitatively predominant variant was detected in the infant. In two of the latter, the predominant variant was mutant; in three it was wild type.
In this analysis, a mutant/wild-type mixed population was detected for a total of eight maternal virus mutations; in each case only one variant was observed in the infant. In five of the eight mixed populations, the transmitted sequence was both wild type and predominant; in two, the transmitted sequence was mutant and predominant; and in only one was it wild type and minority. There were no instances of mutant/minority being transmitted.
There was evidence of emergence of additional resistance mutations in infants. Of 15 mothers with wild-type virus at all resistance-associated codons, 11 had infants with fully concordant virus. The four remaining infants had a single resistance mutation each: M36M/I at 127 days (pair 6), T215T/Y at 42 days (pair 10), L74V/L at 170 days (pair 21), and K70R at 31 days (pair 14).
We have characterized resistance mutations found in virus from 32 HIV-transmitting mothers and their infants. While viral genotypes were not always identical, those of the infants closely matched the mother's virus.
Our previous work examined only zidovudine-associated resistance mutations. The present work extends these analyses to resistance mutations currently seen in the multidrug era. Previously, we had observed that the mutant variant solely was transmitted when maternal virus was mixed at reverse transcriptase codons 210 and 215 but wild type was transmitted in three cases where mutant/wild-type mixtures were present at codon 70 . In the present series, we observed two additional HIV-transmitting mothers with K70K/R mixtures; again both infants evidenced only the wild type despite continued drug pressure with infant zidovudine treatment. The predominance of wild-type sequence in both maternal mixtures suggests transmission may be a result of quantitative advantage, rather than a reduction of in-vivo transmission fitness with K70R mutation.
One patient had a K103N mutation indicative of NNRTI resistance. Since this mutation was observed in a specimen collected 15 days postpartum and use of nevirapine in labor may result in rapid emergence of the K103N mutation , we conducted a chart review to exclude an unrecorded dose. A review of treatment dates confirmed that the mother received no NNRTI drug during pregnancy or delivery, but she had received efavirenz as part of a research protocol prior to pregnancy. The observance of the K103N mutant is then of note, as it was likely present at delivery but not transmitted.
Neither major PI-associated mutation observed (L90M or V82I/V) was transmitted. This is consistent with in-vitro evidence that mutations at these codons result in loss of replicative fitness [19,20], and this may extend to transmission fitness. However, as only two such mutations were present it may also be purely chance.
Most minor PI-associated mutations were transmitted. Since only one mother used a PI during pregnancy, these likely represent naturally occurring polymorphisms, or primary maternal infection from a source treated with PI drugs.
It appears, consistent with our previous report, that where maternal virus is a mixture of mutant and wild type at a specific codon, only one variant is transmitted. For seven of eight maternal mixed viral populations, the variant transmitted was quantitatively predominant. Therefore, quantity may ultimately be more important than replicative quality, and this hypothesis should be subjected to further research. The exception was the PI-associated major mutation discussed above.
It is important to note that while ART-associated mutations may not be preferentially transmitted, they are transmitted. Mothers in this study did transmit multiple ART-associated mutations to their infants, and such transmission could result in reduced treatment options and altered progression of disease in these infants.
This study has limitations. While the sample represents the largest such series of paired mother–infant virus sequences presented to date, it is not large enough in itself to draw firm conclusions from the data. Nevertheless, the descriptive results suggest a need for additional research, and the findings should be subject to further analysis in other study populations with larger sample sizes to permit statistical testing and evaluation.
In addition, it is possible in some cases that mutations detected do not accurately represent those present in patient viral populations, or that concordant mutations result from reasons other than transmission. Since most vertical transmission in non-breastfeeding populations occurs during labor and delivery, it is frequently not possible to isolate HIV-1 from infants at delivery. In this analysis, infant specimens were obtained a median of 39 days after delivery (range, 0–180). In this post-076 era, all infants were treated with ART from birth, allowing the possibility that virus populations detected may have evolved since transmission. Indeed, several new mutations were present in infants, including the only three infant mutant/wild-type mixtures observed. While these new mutations were consistent with infant treatment, they could also represent transmissions of minor maternal variants. A second limitation is that the use of viral isolates obtained from culture allows the possibility of brief continued viral evolution in the absence of drug pressure. Therefore, it is possible that some mutations present in patients' PBMC could have been ‘lost’ in culture and not detected in this analysis. While it is also possible that new mutations could emerge in culture, the sequence results obtained for 18 patients with the methods described above were compared with paired specimens obtained from direct PCR amplification of virus from patient plasma obtained at the same clinic visit (results not shown) and indicate that this is unlikely. Paired sequences demonstrated strong but imperfect concordance, with all differences in a single direction: mutations in virus sequenced from plasma not being detectable in virus obtained from drug-free culture. Therefore, it is likely that where our results err, they err in failure to detect existing mutations rather than in detection of mutations not existing in patient's virus.
The increasing use of ART therapy worldwide will undoubtedly impose selection pressures for HIV-1 viral populations toward continued evolution of resistance. As the prevalence of resistance mutations increases, so more attention should be paid to the transmission characteristics of resistance-associated variants, in vivo as well as in vitro.
Sponsorship: This study is supported by the US National Institutes of Health (NIAID/NIH) grant RO1 AI39144-01. Women and Infants Transmission Study is supported by 1 U01 A150274-01, University of Puerto Rico, San Juan, Puerto Rico (U01 AI 34858), Boston/Worcester Site, Boston, Massachusetts (U01 AI 34856), Columbia Presbyterian Hospital, New York (U01 AI 34842), State University of New York, Brooklyn, New York (HD-3–6117 and RO-1–IID-25714), University of Illinois at Chicago, Chicago, Illinois (U01 AI 34841), Baylor College of Medicine, Houston, Texas (U01 AI 34840), Clinical Trials & Surveys Corp., Baltimore, Maryland (N01 AI 85339).
Note: Dr. Pitt is deceased.
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Women and Infants Transmission Study scientific leadership core: Kenneth Rich (principal investigator) and Delmyra Turpin (study coordinator). Other members include Clemente Diaz and Edna Pacheco-Acosta (University of Puerto Rico, San Juan, Puerto Rico); Ruth Tuomala, Ellen Cooper, and Donna Mesthene (Boston/Worcester Site, Boston); Philip LaRussa and Alice Higgins (Columbia Presbyterian Hospital, New York); Sheldon Landesman, Edward Handelsman, and Gail Moroso (State University of New York, Brooklyn, New York); Kenneth Rich and Delmyra Turpin (University of Illinois at Chicago, Chicago); William Shearer, Susan Pacheco, and Norma Cooper (Baylor College of Medicine, Houston); Joana Rosario (National Institutes of Allergy and Infectious Disease, Bethesda); Robert Nugent, (National Institute of Child Health and Human Development, Bethesda); Vincent Smeriglio and Katherine Davenny (National Institute on Drug Abuse, Rockville); and Bruce Thompson (Clinical Trials & Surveys Corp., Baltimore).
20. Martinez-Picado J, Savara AV, Shi L, Sutton L, D'Aquila RT. Fitness of human immunodeficiency virus type 1 protease inhibitor-selected single mutants. Virology 2000; 275:318–322.
Keywords:© 2006 Lippincott Williams & Wilkins, Inc.
HIV; anti-HIV drugs; viral RNA; vertical transmission; drug resistance