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Human Mammary Tumor Virus (HMTV) sequences in human milk.

Nartey T, Moran H, Marin T, Arcaro KF, Anderton DL, Etkind P, Holland JF, Melana SM, Pogo BG - Infect. Agents Cancer (2014)

Bottom Line: This virus was designated as human mammary tumor virus (HMTV).In the remaining 65 women of the Biopsy-Group, under enough clinical suspicion to lead to biopsy, HMTV was detected in 14, nearly three times the number of milks as compared to the Reference-Group (21.54% versus 7.61%; p: 0.016).The similarity of HMTV to MMTV is striking and suggests one possible avenue for viral transmission in humans.

View Article: PubMed Central - HTML - PubMed

Affiliation: The Tisch Cancer Institute and Department of Medicine, Icahn School of Medicine at Mount Sinai, 1 Gustave L. Levy Place, New York, NY 10029, USA.

ABSTRACT

Background: Retroviral sequences 90-95% homologous to the mouse mammary tumor virus (MMTV) were present in 38% of the breast cancers studied from American women and were not detectable in non-tumor breast tissue from the same patient. The entire proviral structure was described and viral particles were isolated from primary cultures of human breast cancer. This virus was designated as human mammary tumor virus (HMTV). Hormone response elements present in the HMTV Long-Terminal-Repeat (LTR) suggest a mechanism for association of HMTV with hormonally responding tissues. In fact, the incidence of HMTV sequences is higher in gestational breast cancers, which are associated with hormonal changes. Milk epithelial cells are also under hormonal regulation and therefore are excellent specimens for HMTV sequence detection.

Methods: The HMTV sequence was studied in milk samples from lactating women recruited with increased risk of breast cancer because they had undergone breast biopsies (Biopsy-Group) and lactating women without breast biopsies (Reference-Group).

Results: HMTV-env sequences were detected by PCR in milk of 7.61% of 92 women of the Reference-Group and in 20.55% of 73 women of the Biopsy-Group (p: 0.015). The sequences were 94-98% homologous to MMTV. HMTV-env and HMTV-env/LTR junction sequences were detected in high-speed pellet RNA, implying the presence of HMTV viral particles. PCR assays to detect the murine mitochondrial cytochrome oxidase gene and intracisternal-A-type particle sequences were performed to rule out mouse mitochondrial or genomic DNA contamination. Eight women of the 73 Biopsy-Group participants had breast cancer and the milk of only one of these eight women had HMTV-env sequences. In the remaining 65 women of the Biopsy-Group, under enough clinical suspicion to lead to biopsy, HMTV was detected in 14, nearly three times the number of milks as compared to the Reference-Group (21.54% versus 7.61%; p: 0.016).

Conclusion: The significance of HMTV in milk from the Reference-Group, the greater frequency in the milk of women who had undergone a breast biopsy and its possible infectivity for infants are important questions under study. The similarity of HMTV to MMTV is striking and suggests one possible avenue for viral transmission in humans.

No MeSH data available.


Related in: MedlinePlus

Detection of HMTV-env gene sequences in P2 RNA. cDNA synthesized from high speed pellet (P2) RNA using polydT as primer, PCR was performed using primers 5 L and 3 L and nested PCR using primers 1XXX and 3 F as described in Methods. A. Diagram of HMTV viral RNA structure and location of the primers. B. HMTV-env cDNA sequence was amplified using primers 1XXX and 3 F in a nested PCR reaction (GenBank KJ831810). C. 2% agarose gel electrophoresis of HMTV-env 251 bp amplicon. Lane 1: HMTV-env positive sample. Lane 2: 1 kb plus DNA ladder. Lane 3: Southern blot hybridization using 32P end labeled probe for HMTV-env positive sample.
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Figure 2: Detection of HMTV-env gene sequences in P2 RNA. cDNA synthesized from high speed pellet (P2) RNA using polydT as primer, PCR was performed using primers 5 L and 3 L and nested PCR using primers 1XXX and 3 F as described in Methods. A. Diagram of HMTV viral RNA structure and location of the primers. B. HMTV-env cDNA sequence was amplified using primers 1XXX and 3 F in a nested PCR reaction (GenBank KJ831810). C. 2% agarose gel electrophoresis of HMTV-env 251 bp amplicon. Lane 1: HMTV-env positive sample. Lane 2: 1 kb plus DNA ladder. Lane 3: Southern blot hybridization using 32P end labeled probe for HMTV-env positive sample.

Mentions: HMTV particle sequences were also sought in the high-speed pellet (P2) RNA from milk of the same Biopsy and Reference Group women whose milk cell DNA had HMTV sequences. RNA was extracted as described in Methods and RT-PCR carried out for HMTV-env (Figure 2) and HMTV-env/LTR junction sequences (Figure 3). The HMTV sequences from the cDNA were compared to sequences present in the GenBank. HMTV-env sequences were 94 to 98% homologous to MMTV (GenBank: M15122) and HMTV-env/LTR junction sequence was 96% homologous to MMTV (GenBank: M15122). Results in Table 2 indicate that HMTV-env and HMTV-env/LTR junction sequences were amplified from P2 RNA when the HMTV-env also was detected in the genomic DNAs from P1. HMTV-env sequences were amplified in P2 RNA from 2 out of 3 women of the Reference-Group. All three of these women also had HMTV sequences detected in their genomic DNA and HMTV-env/LTR junction sequences present in P2 RNA. In the case of the Biopsied-Group, HMTV-env and HMTV env/LTR junction sequences were amplified in P2 RNA from 4 out of 5 women whose genomic DNA contained HMTV sequences. HMTV-env/LTR junction sequences were detected in a total of 7 P2 RNA samples of which one contained enough material to allow sequencing. HMTV-env and env/LTR junction sequences from P2 RNA have been submitted to the GenBank. Absence of glycerol-3-phosphate dehydrogenase gene (G3DPH) mRNA from high-speed pellets and the presence of HMTV-env/LTR junction sequences is consistent with the presence of virion RNA.


Human Mammary Tumor Virus (HMTV) sequences in human milk.

Nartey T, Moran H, Marin T, Arcaro KF, Anderton DL, Etkind P, Holland JF, Melana SM, Pogo BG - Infect. Agents Cancer (2014)

Detection of HMTV-env gene sequences in P2 RNA. cDNA synthesized from high speed pellet (P2) RNA using polydT as primer, PCR was performed using primers 5 L and 3 L and nested PCR using primers 1XXX and 3 F as described in Methods. A. Diagram of HMTV viral RNA structure and location of the primers. B. HMTV-env cDNA sequence was amplified using primers 1XXX and 3 F in a nested PCR reaction (GenBank KJ831810). C. 2% agarose gel electrophoresis of HMTV-env 251 bp amplicon. Lane 1: HMTV-env positive sample. Lane 2: 1 kb plus DNA ladder. Lane 3: Southern blot hybridization using 32P end labeled probe for HMTV-env positive sample.
© Copyright Policy - open-access
Related In: Results  -  Collection

License 1 - License 2
Show All Figures
getmorefigures.php?uid=PMC4129428&req=5

Figure 2: Detection of HMTV-env gene sequences in P2 RNA. cDNA synthesized from high speed pellet (P2) RNA using polydT as primer, PCR was performed using primers 5 L and 3 L and nested PCR using primers 1XXX and 3 F as described in Methods. A. Diagram of HMTV viral RNA structure and location of the primers. B. HMTV-env cDNA sequence was amplified using primers 1XXX and 3 F in a nested PCR reaction (GenBank KJ831810). C. 2% agarose gel electrophoresis of HMTV-env 251 bp amplicon. Lane 1: HMTV-env positive sample. Lane 2: 1 kb plus DNA ladder. Lane 3: Southern blot hybridization using 32P end labeled probe for HMTV-env positive sample.
Mentions: HMTV particle sequences were also sought in the high-speed pellet (P2) RNA from milk of the same Biopsy and Reference Group women whose milk cell DNA had HMTV sequences. RNA was extracted as described in Methods and RT-PCR carried out for HMTV-env (Figure 2) and HMTV-env/LTR junction sequences (Figure 3). The HMTV sequences from the cDNA were compared to sequences present in the GenBank. HMTV-env sequences were 94 to 98% homologous to MMTV (GenBank: M15122) and HMTV-env/LTR junction sequence was 96% homologous to MMTV (GenBank: M15122). Results in Table 2 indicate that HMTV-env and HMTV-env/LTR junction sequences were amplified from P2 RNA when the HMTV-env also was detected in the genomic DNAs from P1. HMTV-env sequences were amplified in P2 RNA from 2 out of 3 women of the Reference-Group. All three of these women also had HMTV sequences detected in their genomic DNA and HMTV-env/LTR junction sequences present in P2 RNA. In the case of the Biopsied-Group, HMTV-env and HMTV env/LTR junction sequences were amplified in P2 RNA from 4 out of 5 women whose genomic DNA contained HMTV sequences. HMTV-env/LTR junction sequences were detected in a total of 7 P2 RNA samples of which one contained enough material to allow sequencing. HMTV-env and env/LTR junction sequences from P2 RNA have been submitted to the GenBank. Absence of glycerol-3-phosphate dehydrogenase gene (G3DPH) mRNA from high-speed pellets and the presence of HMTV-env/LTR junction sequences is consistent with the presence of virion RNA.

Bottom Line: This virus was designated as human mammary tumor virus (HMTV).In the remaining 65 women of the Biopsy-Group, under enough clinical suspicion to lead to biopsy, HMTV was detected in 14, nearly three times the number of milks as compared to the Reference-Group (21.54% versus 7.61%; p: 0.016).The similarity of HMTV to MMTV is striking and suggests one possible avenue for viral transmission in humans.

View Article: PubMed Central - HTML - PubMed

Affiliation: The Tisch Cancer Institute and Department of Medicine, Icahn School of Medicine at Mount Sinai, 1 Gustave L. Levy Place, New York, NY 10029, USA.

ABSTRACT

Background: Retroviral sequences 90-95% homologous to the mouse mammary tumor virus (MMTV) were present in 38% of the breast cancers studied from American women and were not detectable in non-tumor breast tissue from the same patient. The entire proviral structure was described and viral particles were isolated from primary cultures of human breast cancer. This virus was designated as human mammary tumor virus (HMTV). Hormone response elements present in the HMTV Long-Terminal-Repeat (LTR) suggest a mechanism for association of HMTV with hormonally responding tissues. In fact, the incidence of HMTV sequences is higher in gestational breast cancers, which are associated with hormonal changes. Milk epithelial cells are also under hormonal regulation and therefore are excellent specimens for HMTV sequence detection.

Methods: The HMTV sequence was studied in milk samples from lactating women recruited with increased risk of breast cancer because they had undergone breast biopsies (Biopsy-Group) and lactating women without breast biopsies (Reference-Group).

Results: HMTV-env sequences were detected by PCR in milk of 7.61% of 92 women of the Reference-Group and in 20.55% of 73 women of the Biopsy-Group (p: 0.015). The sequences were 94-98% homologous to MMTV. HMTV-env and HMTV-env/LTR junction sequences were detected in high-speed pellet RNA, implying the presence of HMTV viral particles. PCR assays to detect the murine mitochondrial cytochrome oxidase gene and intracisternal-A-type particle sequences were performed to rule out mouse mitochondrial or genomic DNA contamination. Eight women of the 73 Biopsy-Group participants had breast cancer and the milk of only one of these eight women had HMTV-env sequences. In the remaining 65 women of the Biopsy-Group, under enough clinical suspicion to lead to biopsy, HMTV was detected in 14, nearly three times the number of milks as compared to the Reference-Group (21.54% versus 7.61%; p: 0.016).

Conclusion: The significance of HMTV in milk from the Reference-Group, the greater frequency in the milk of women who had undergone a breast biopsy and its possible infectivity for infants are important questions under study. The similarity of HMTV to MMTV is striking and suggests one possible avenue for viral transmission in humans.

No MeSH data available.


Related in: MedlinePlus