Effects of hepatitis B virus infection on human sperm chromosomes.
OBJECTIVE
To evaluate the level of sperm chromosome aberrations in male patients with hepatitis B, and to directly detect whether there are HBV DNA integrations in sperm chromosomes of hepatitis B patients.


METHODS
Sperm chromosomes of 14 tested subjects (5 healthy controls, 9 patients with HBV infection, including 1 with acute hepatitis B, 2 with chronic active hepatitis B, 4 with chronic persistent hepatitis B, 2 chronic HBsAg carriers with no clinical symptoms) were prepared using interspecific in vitro fertilization between zona-free golden hamster ova and human spermatozoa, and the frequencies of aberration spermatozoa were compared between subjects of HBV infection and controls. Fluorescence in situ hybridization (FISH) to sperm chromosome spreads was carried out with biotin-labeled full length HBV DNA probe to detect the specific HBV DNA sequences in the sperm chromosomes.


RESULTS
The total frequency of sperm chromosome aberrations in HBV infection group (14.8 %, 33/223) was significantly higher than that in the control group (4.3 %, 5/116). Moreover, the sperm chromosomes in HBV infection patients commonly presented stickiness, clumping, failure to staining, etc, which would affect the analysis of sperm chromosomes. Specific fluorescent signal spots for HBV DNA were seen in sperm chromosomes of one patient with chronic persistent hepatitis. In 9 (9/42) sperm chromosome complements containing fluorescent signal spots, one presented 5 obvious FISH spots, others presented 2 to 4 signals. There was significant difference of fluorescence intensity among the signal spots. The distribution of signal sites among chromosomes was random.


CONCLUSIONS
HBV infection can bring about mutagenic effects on sperm chromosomes. Integrations of viral DNA into sperm chromosomes which are multisites and nonspecific, can further increase the instability of sperm chromosomes. This study suggested that HBV infection can create extensively hereditary effects by alteration genetic constituent and/or induction chromosome aberrations, as well as the possibility of vertical transmission of HBV via the germ line to the next generation.
Huang
Jian-Min
JM
Department of Cell Biology and Medical Genetics, Shantou University Medical College, Shantou 515031, Guangdong Province, China.
Huang
Tian-Hua
TH
Qiu
Huan-Ying
HY
Fang
Xiao-Wu
XW
Zhuang
Tian-Gang
TG
Liu
Hong-Xi
HX
Wang
Yong-Hua
YH
Deng
Li-Zhi
LZ
Qiu
Jie-Wen
JW
D000328:Adult; D002877:Chromosomes, Human / Q000235:genetics*; D004279:DNA, Viral / Q000032:analysis; D006511:Hepatitis B Antigens / Q000032:analysis / Q000097:blood; D006514:Hepatitis B Surface Antigens / Q000032:analysis / Q000097:blood; D019694:Hepatitis B, Chronic / Q000235:genetics*; D006801:Humans; D017404:In Situ Hybridization, Fluorescence; D007621:Karyotyping; D008297:Male; D012016:Reference Values; D012661:Semen / Q000276:immunology; D013094:Spermatozoa / Q000821:virology*
D004279:DNA, Viral; D006511:Hepatitis B Antigens; D006514:Hepatitis B Surface Antigens