Abstract
We used a nested polymerase chain reaction (PCR) to diagnose HTLV‐1 carriers. The DNA isolated from the nuclear extract obtained from frozen whole blood was found appropriate for PCR study both qualitatively and quantitatively. The use of freshly frozen whole blood made the field work much easier, and the use of a nuclear extraction procedure allowed DNA isolation in just 4 microcentrifuge tubes. We could not attain sufficient sensitivity to detect a single molecule with single‐step PCR, but nested PCR was confirmed to detect a single molecule/reaction. All samples of the seropositive group including 94 blood donors, 66 mothers, and 13 children were positive in the nested PCR, while none of the seronegative group, including 198 blood donors and 285 children, was positive. Although 18/717 (2.5%) cord blood samples obtained from babies born to carrier mothers were PCR‐positive, none of 5 formula‐fed children tested who had been PCR‐positive in the cord blood gave evidence of infection later on. Furthermore, all of 4 seropositive infected children who were formula‐fed had been PCR‐negative in their cord blood. The results are not consistent with intrauterine infection, but suggest the presence of a perinatal or postnatal infection route other than through breast milk.
Keywords: Key words, Cord blood, HTLV‐1, Maternal transmission, Milk‐borne infection, PCR
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REFERENCES
- 1. ) Poiesz , B.J. , Ruscetti , F. W. , Gazdar , A. F. , Bunn , P. A. , Minna , J. D. and Gallo , R. C.Detection and isolation of type C retro virus particles from fresh and cultured lymphocytes of a patient with cutaneous T‐cell lymphoma . Proc. Natl. Acad. Sci. USA , 77 , 7415 – 7419 ( 1980. ). [DOI] [PMC free article] [PubMed] [Google Scholar]
- 2. ) Hinuma , Y. , Nagata , K. , Hanaoka , M.Nakai , M. , Matsumoto , T. , Kinoshita , K. , Shirakawa , S. and Miyoshi , I.Adult T‐cell leukemia: antigen in an ATL cell line and 975 detection of antibodies to the antigen in human sera . Proc. Natl. Acad. Sci USA , 78 , 6476 – 6480 ( 1981. ). [DOI] [PMC free article] [PubMed] [Google Scholar]
- 3. ) Yoshida , M. , Miyoshi , I. and Hinuma , Y.Isolation and characterization of retrovirus from cell lines of human adult T‐cell leukemia and its implication in the disease . Proc. Natl. Acad. Sci USA , 79 , 2031 – 2035 ( 1982. ). [DOI] [PMC free article] [PubMed] [Google Scholar]
- 4. ) Uchiyama , T. , Yodoi , J. , Sagawa , K. , Takatsuki , K. and Uchino , H.Adult T‐cell leukemia: clinical and hematologic features of 16 cases . Blood , 50 , 481 – 492 ( 1977. ). [PubMed] [Google Scholar]
- 5. ) Yoshida , M. , Seiki , M. , Yamaguchi , K. and Takatsuki , K.Monoclonal integration of human T‐cell leukemia provirus in all primary tumors of adult T‐cell leukemia suggests causative role of human T‐cell leukemia virus in the disease . Proc. Natl. Acad. Sci USA , 81 , 2534 – 2537 ( 1984. ). [DOI] [PMC free article] [PubMed] [Google Scholar]
- 6. ) Gessain , A. , Barin , F. , Vernant , J. C. , Gout , O. , Maurs , L. and Calender , A.Antibodies to human T‐lymphotropic virus type‐1 in patients with tropical spastic paraparesis . Lancet , ii , 407 – 410 ( 1985. ). [DOI] [PubMed] [Google Scholar]
- 7. ) Osame , M. , Usuku , K. , Izumo , S. , Ijichi , N. , Amitani , H. , Igata , A. , Matsumoto , M. and Tara , M.HTLV‐1 associated myelopathy, a new clinical entity . Lancet , i , 1031 – 1032 ( 1986. ). [DOI] [PubMed] [Google Scholar]
- 8. ) Hinuma , Y. , Komoda , H. , Chosa , T. , Kondo , T. , Kohakura , M. , Takenaka , T. , Kikuchi , M. , Ichimaru , M. , Yunoki , K. , Sato , I. , Matsuo , R. , Takiuchi , Y. , Uchino , H. and Hanaoka , M.Antibodies to adult T‐cell leukemia‐virus‐associated antigen (ATLA) in sera from patients with ATL and controls in Japan: a nation‐wide sero‐epidemiologic study . Int. J. Cancer , 29 , 631 – 635 ( 1982. ). [DOI] [PubMed] [Google Scholar]
- 9. ) The T‐ and B‐cell malignancy study group . The third nation‐wide study on adult T‐cell leukemia/lymphoma (ATL) in Japan: characteristic patterns of HLA antigen and HTLV‐1 infection in ATL patients and their relatives . Int. J. Cancer , 41 , 505 – 512 ( 1988. ). [DOI] [PubMed] [Google Scholar]
- 10. ) Blattner , W. A. , Kalyanaraman , V. S. , Robert‐Guroff , ML , Lister , T. A. , Galton , D. A. G. , Sarin , P. S. , Crawford , M. H. , Catovsky , D. , Greaves , M. and Gallo , R. C.The human type‐C retrovirus, HTLV, in blacks from the Caribbean region, and relationship to adult T‐cell leukemia/ lymphoma . Int. J. Cancer , 30 , 257 – 264 ( 1982. ). [DOI] [PubMed] [Google Scholar]
- 11. ) Fleming , A. F. , Yamamoto , N. , Bhusnurmath , S. R. , Maharajan , R. , Schneider , J. and Hunsmann , G.Antibodies to ATLV (HTLV) in Nigerian blood donors and patients with chronic lymphatic leukaemia or lymphoma . Lancet , ii , 334 – 335 ( 1983. ). [DOI] [PubMed] [Google Scholar]
- 12. ) Tajima , K. , Tominaga , S. , Suchi , T. , Kawagoe , T. , Komoda , H. , Hinuma , Y. , Oda , T. and Fujita , K.Epidemiological analysis of the distribution of antibody to adult T‐cell leukemia‐virus‐associated antigen: possible horizontal transmission of adult T‐cell leukemia virus . Gann , 73 , 893 – 901 ( 1982. ). [PubMed] [Google Scholar]
- 13. ) Hino , S. , Yamaguchi , K. , Katamine , S. , Sugiyama , H. , Amagasaki , T. , Kinoshita , K. , Yoshida , Y. , Doi , H. , Tsuji , Y. and Miyamoto , T.Mother‐to‐child transmission of human T‐cell leukemia virus type‐I . Jpn. J. Cancer Res. , 76 , 474 – 480 ( 1985. ). [PubMed] [Google Scholar]
- 14. ) Kinoshita , K. , Kamihira , S. , Yamada , Y. , Amagasaki , T. , Ikeda , S. , Momita , S. and Ichimaru , M.Adult T cell leukemia‐lymphoma in the Nagasaki district . Gann Monogr. Cancer Res. , 28 , 167 – 184 ( 1982. ). [Google Scholar]
- 15. ) Kaplan , J. E. , Osame , M. , Kubota , H. , Igata , A. , Nishitani , H. , Maeda , Y. , Khabbaz , R. F. and Janssen , R. S.The risk of development of HTLV‐I‐associated myelopathy/tropical spastic paraparesis among persons infected with HTLV‐I . J. Acquired Immune Defic. Syndr. , 3 , 1096 – 1101 ( 1990. ). [PubMed] [Google Scholar]
- 16. ) Hino , S. , Sugiyama , H. , Doi , H. , Ishimaru , T. , Yamabe , T. , Tsuji , Y. and Miyamoto , T.Breaking the cycle of HTLV‐1 transmission via carrier mothers' milk . Lancet , ii , 158 – 159 ( 1987. ). [DOI] [PubMed] [Google Scholar]
- 17. ) Hino , S.Maternal‐infant transmission of HTLV‐1: implication for disease . In “ Human Retrovirology: HTLV ,” ed. Blattner W. A. , pp. 363 – 375 ( 1990. ). Raven Press; , New York . [Google Scholar]
- 18. ) Gotoh , Y. I , Sugamura , K. and Hinuma , Y.Healthy carriers of human retrovirus, adult T‐cell leukemia virus (ATLV): demonstration by clonal culture of ATLV‐carrying T cells from peripheral blood . Proc. Natl. Acad, Sci. USA , 79 , 4780 – 4782 ( 1982. ). [DOI] [PMC free article] [PubMed] [Google Scholar]
- 19. ) Saito , S. , Furuki , K. , Ando , Y. , Tanigawa , T. , Kakimoto , K. , Moriyama , I. and Ichijo , M.Identification of HTLV‐I sequence in cord blood mononuclear cells of neonates born to HTLV‐I antigen/antibody‐positive mothers by polymerase chain reaction . Jpn. J. Cancer Res. , 81 , 890 – 895 ( 1990. ). [DOI] [PMC free article] [PubMed] [Google Scholar]
- 20. ) Abbott , M. A. , Poiesz , B. J. , Byrne , B. C. , Kwok , S. , Sninsky , J. J. and Ehrlich , G. D.Enzymatic gene amplification: qualitative and quantitative methods for detecting proviral DNA amplified in vitro . J. Infect. Dis. , 158 , 1158 – 1169 ( 1988. ). [DOI] [PubMed] [Google Scholar]
- 21. ) Fujino , R. , Kawato , K. , Ikeda , M. , Miyakoshi , H. , Mizukoshi , M. and Imai , J.Improvement of gelatin particle agglutination test for detection of anti‐HTLV‐I antibody . Jpn. J. Cancer Res. , 82 , 367 – 370 ( 1991. ). [DOI] [PMC free article] [PubMed] [Google Scholar]
- 22. ) Ikeda , M. , Fujino , R. , Matsui , T. , Yoshida , T. , Komoda , H. and Imai , J.A new agglutination test for serum antibodies to adult T‐cell leukemia virus . Gann , 75 , 845 – 848 ( 1984. ). [PubMed] [Google Scholar]
- 23. ) Nam , S.H. and Hatanaka , M.Identification of a protease gene of human T‐cell leukemia virus type I (HTLV‐I) and its structural comparison . Biochem. Biophys. Res. Commun. , 139 , 129 – 135 ( 1986. ). [DOI] [PubMed] [Google Scholar]
- 24. ) Foley , G. E. , Lazarus , H. , Farber , S. , Uzman , B. G. , Boone , B. A. and McCarthy , R. E.Continuous culture of human lymphoblasts from peripheral blood of a child with acute leukemia . Cancer , 18 , 522 – 529 ( 1965. ). [DOI] [PubMed] [Google Scholar]
- 25. ) Maeda , Y. , Imai , J. , Kiyokawa , H. , Kanamura , M. and Hino , S.Performance certification of gelatin particle agglutination assay for anti‐HTLV‐1 antibody: inconclusive positive results . Jpn. J. Cancer Res. , 80 , 915 – 919 ( 1989. ). [DOI] [PMC free article] [PubMed] [Google Scholar]
- 26. ) Beutler , E. , Gelbart , T. and Kuhl , W.Interference of heparin with polymerase chain reaction . Bio Techniques , 9 , 166 ( 1990. ). [PubMed] [Google Scholar]
- 27. ) Clarke , M. , F. , Gelman , E. P. and Reitz , M. S. , Jr.Homology of human T‐cell leukaemia virus envelope gene with class I HLA gene . Nature , 305 , 60 – 62 ( 1983. ). [DOI] [PubMed] [Google Scholar]
- 28. ) Katsuki , T. and Hinuma , Y.Characteristics of cell lines derived from human leukocytes transformed by different strains of Epstein‐Barr virus . Int. J. Cancer , 15 , 203 – 210 ( 1975. ). [DOI] [PubMed] [Google Scholar]
- 29. ) Maniatis , T. , Fritsch , E. F. and Sambrook , J. “ Molecular Cloning: A Laboratory Manual ” ( 1982. ). Cold Spring Harbor Laboratory; , New York . [Google Scholar]
- 30. ) Higuchi , R.Rapid, efficient DNA extraction for PCR from cells or blood . Amplifications, a Forum for PCR Users (Perkin Elmer Cetus Co.) , 2 , 1 – 3 ( 1989. ). [Google Scholar]
- 31. ) Seiki , M. , Hattori , S. , Hirayama , Y. and Yoshida , M.Human adult T‐cell leukemia virus: complete nucleotide sequence of the provirus genome integrated in leukemia cell DNA . Proc. Natl Acad. Sci. USA , 80 , 3618 – 3622 ( 1983. ). [DOI] [PMC free article] [PubMed] [Google Scholar]
- 32. ) Bonthron , D. T. , Morton , C. C. , Orkin , S. H. and Collins , T.Platelet‐derived growth factor A chain: gene structure, chromosomal location, and basis for alternative mRNA splicing . Proc. Natl. Acad. Sci. USA , 85 , 1492 – 1496 ( 1988. ). [DOI] [PMC free article] [PubMed] [Google Scholar]
- 33. ) Saiki , R. K. , Scharf , S. , Faloona , F. , Mullis , K. B. , Horn , G. T. , Erlich , H. A. and Arnheim , N.Enzymatic amplification of β‐globin genomic sequences and restriction site analysis for diagnosis of sickle cell anemia . Science , 230 , 1350 – 1354 ( 1985. ). [DOI] [PubMed] [Google Scholar]
- 34. ) Smit , V. T. H. B. M. , Boot , A. J. M. , Smit , A. M. M. , Fleuren , G. J. , Cornelisse , C. J. and Bos , J. L.K‐RAS codon 12 mutations occur very frequently in pancreatic adenocarcinomas . Nucleic Acids Res. , 16 , 7773 – 7782 ( 1988. ). [DOI] [PMC free article] [PubMed] [Google Scholar]
- 35. ) Matsumoto , C. , Mitsunaga , S. , Oguchi , T. , Mitomi , Y. , Shimada , T. , Ichikawa , A. , Watanabe , J , and Nishioka , K.Detection of human T‐cell leukemia virus type 1 (HTLV‐1) provirus in an infected cell line and in peripheral mononuclear cells of blood donors by the nested double polymerase chain reaction method: comparison with HTLV‐1 antibody tests . J. Virol , 64 , 5290 – 5294 ( 1990. ). [DOI] [PMC free article] [PubMed] [Google Scholar]
- 36. ) Ando , Y. , Nakano , S. , Saito , K. , Shimamoto , I. , Ichijo , M. , Toyama , T. and Hinuma , Y.Transmission of adult T‐cell leukemia retro virus (HTLV‐I) from mother to child: comparison of bottle‐ with breast‐fed babies . Jpn. J. Cancer Res. , 78 , 322 – 324 ( 1987. ). [PubMed] [Google Scholar]
- 37. ) Komuro , A. , Hayami , M. , Fujii , H. , Miyahara , S. and Hirayama , M.Vertical transmission of adult T‐cell leukemia virus . Lancet , i , 240 ( 1983. ). [DOI] [PubMed] [Google Scholar]
- 38. ) Satow , Y. , Hashido , M. , Ishikawa , K. , Honda , H. , Mizuno , M. , Kawana , T. and Hayami , M.Detection of HTLV‐1 antigen in peripheral and cord blood lymphocytes from carrier mothers . Lancet , 338 , 915 – 916 ( 1991. ). [DOI] [PubMed] [Google Scholar]
- 39. ) Sugiyama , H. , Doi , H. , Yamaguchi , K. , Tsuji , Y. , Miyamoto , T. and Hino , S.Significance of postnatal mother‐to‐child transmission of human T‐lymphotropic virus type‐1 on the development of adult T‐cell leukemia/lymphoma . J. Med. Virol , 20 , 253 – 260 ( 1986. ). [DOI] [PubMed] [Google Scholar]
- 40. ) Watanabe , T. , Yamaguchi , K. , Takatsuki , K. , Osame , M. and Yoshida , M.Constitutive expression of parathyroid hormone‐related protein gene in human T cell leukemia virus type 1 (HTLV‐1) carriers and adult T cell leukemia patients that can be trans‐activated by HTLV‐1 tax gene . J. Exp. Med. , 172 , 759 – 765 ( 1990. ). [DOI] [PMC free article] [PubMed] [Google Scholar]
- 41. ) Simmonds , P. , Balfe , P. , Peutherer , J. F. , Ludlam , C. A. , Bishop , J. O. and Leigh‐Brown , A. J.Human immunodeficiency virus‐infected individuals contain provirus in small numbers of peripheral mononuclear cells and at low copy numbers . J. Virol , 64 , 864 – 872 ( 1990. ). [DOI] [PMC free article] [PubMed] [Google Scholar]
- 42. ) Kusuhara , K. , Sonoda , S. , Takahashi , K. , Tokugawa , K. , Fukushige , J. and Ueda , K.Mother‐to‐child transmission of human T‐cell leukemia virus type I (HTLV‐I): a fifteen‐year follow‐up study in Okinawa, Japan . Int. J. Cancer , 40 , 755 – 757 ( 1987. ). [DOI] [PubMed] [Google Scholar]
- 43. ) Pate , E. J. , Wiktor , S. Z. , Shaw , G. M. , Champegnie , E. , Murphy , E. L. and Blattner , W. A.Lack of viral latency of human T‐cell lymphotropic virus type 1 . N. Engl. J. Med. , 325 , 284 ( 1991. ). [DOI] [PubMed] [Google Scholar]
- 44. ) Nakano , S. , Ando , Y. , Saito , K. , Moriyama , I. , Ichijo , M. , Toyama , T. , Sugamura , K. , Imai , J. and Hinuma , Y.Primary infection of Japanese infants with adult T‐cell leukemia‐associated retrovirus (ATLV): evidence for viral transmission from mothers to children . J. Infect. , 12 , 205 – 212 ( 1986. ). [DOI] [PubMed] [Google Scholar]
- 45. ) Sugamura , K. , Fujii , M. , Ueda , S. and Hinuma , Y.Identification of a glycoprotein, gp21, of adult T cell leukemia virus by monoclonal antibody . J. Immunol , 132 , 3180 – 3184 ( 1984. ). [PubMed] [Google Scholar]
- 46. ) Shiraishi , Y. , Fujishita , M. , Miyoshi , I. , Kondo , N. and Orii , T.Transformation of Bloom syndrome T‐lymphocytes by co‐cultivation with a lethally irradiated human T‐cell line carrying type C virus particles . Gann , 74 , 213 – 218 ( 1983. ). [PubMed] [Google Scholar]
- 47. ) Takehara , N. , Iwahara , Y. , Uemura , Y. , Sawada , T. , Ohtsuki , Y. , Iwai , H. , Hoshino , H. and Miyoshi , I.Effect of immunization on HTLV‐1 infection in rabbits . Int. J. Cancer , 44 , 332 – 336 ( 1989. ). [DOI] [PubMed] [Google Scholar]