抗变形链球菌鸡卵黄抗体预防龋齿的动物实验研究
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摘要
目的：制备抗变形链球菌鸡卵黄抗体的粗提物，并研究其对大鼠龋齿
的预防作用。
方法：使用变形链球菌和远缘链球菌加佐剂免疫母鸡，用水稀释——
超滤法从鸡卵黄中提取抗变形链球菌鸡卵黄抗体的粗提物。采
用Lowray's法测定其蛋白含量，用间接血凝实验测定其效价。
S-D大鼠40只建立龋齿模型并以随机区组的方法分为四组。
第一组在饮水中加2％抗变形链球菌鸡卵黄抗体，第二组加
1ppm氟化钠，第三组加2％从未免疫母鸡鸡卵黄中提取的非特
异性鸡卵黄抗体，第四组加蒸馏水。55天后采用keyes方法
对龋齿进行记分。
结果：经免疫后在鸡卵黄中产生了抗变形链球菌鸡卵黄抗体。用水稀
释——超滤法提取的抗变形链球菌鸡卵黄抗体蛋白含量约为
100mg/ml，效价1：512。Keyes龋齿记分经统计分析第一组显
著低于第三组、第四组（P＜0.01），第二组也显著低于第三组、
第四组（P＜0.01）。第一组与第二组无显著差异（P＞0.05）。
结论：用本实验方法制备的抗变形链球菌鸡卵黄抗体能有效的预防大
鼠龋齿的发生，与氟水无显著差异。
Objective: The primary extract of anti-streptococcus nzulans
IgY was prepared and observed it-s pretective effect
on rats-dental caries
The anti-ultreptococcus znulans IgY was primary
extracted from the eggs laid by the hens immunized by
&rnutans and S. sobrinus with adjuvant by water
dilution ultrafiltration method.The protein
content of the anti-ultreptococcus inut~zns IgY was
determined by Lowray's method. The titer of the
anti-ultreptococcus rniitans IgY was examined by Passive
JIemagglutination Assay(PIIA). 40 S-D rats have been
established dental caries model were divided into
four groups with random block degisn: the first group
was given 2% the anti-ultreptococcus nzutans IgY in
daily drinks; the second group was given lpptn sodium
fluoride; the third group was given the non-special IgY
extracted by the same method from the eggs laid by the
hens without iniinunity ; the fourth group was given
4
distilled water. After 55 days of treatment, the caries
scores were measured by the keyes's method.
Results: The anti-ultreptococcus fihlltans IgY was produced in eggs
laid by hens after immunity. The protein content of
primary extract of anti-ultreptococcus nzutans IgY
extracted from eggs yolk by water
dilution ultrafiltration method was 100mg/ml. The
titer of this TgY was 1:512. The keyes' caries score of
the first group was significant lower than the one of
the third group and the fourth group(p<0.01) . The
scores of the second group was significant lower than
it of the third group and the fourth group (p<0.01) too.
There wasn't significant difference between the scores
of the first group and the second group.
Conclusion:The primary extract of anti-ultrpotococcus znutans
JgY extracted by the method of this study could
effective prctected rats against dental caries.The
effect was simi larly with sodium fiuorid.
的预防作用。
方法：使用变形链球菌和远缘链球菌加佐剂免疫母鸡，用水稀释——
超滤法从鸡卵黄中提取抗变形链球菌鸡卵黄抗体的粗提物。采
用Lowray's法测定其蛋白含量，用间接血凝实验测定其效价。
S-D大鼠40只建立龋齿模型并以随机区组的方法分为四组。
第一组在饮水中加2％抗变形链球菌鸡卵黄抗体，第二组加
1ppm氟化钠，第三组加2％从未免疫母鸡鸡卵黄中提取的非特
异性鸡卵黄抗体，第四组加蒸馏水。55天后采用keyes方法
对龋齿进行记分。
结果：经免疫后在鸡卵黄中产生了抗变形链球菌鸡卵黄抗体。用水稀
释——超滤法提取的抗变形链球菌鸡卵黄抗体蛋白含量约为
100mg/ml，效价1：512。Keyes龋齿记分经统计分析第一组显
著低于第三组、第四组（P＜0.01），第二组也显著低于第三组、
第四组（P＜0.01）。第一组与第二组无显著差异（P＞0.05）。
结论：用本实验方法制备的抗变形链球菌鸡卵黄抗体能有效的预防大
鼠龋齿的发生，与氟水无显著差异。
Objective: The primary extract of anti-streptococcus nzulans
IgY was prepared and observed it-s pretective effect
on rats-dental caries
The anti-ultreptococcus znulans IgY was primary
extracted from the eggs laid by the hens immunized by
&rnutans and S. sobrinus with adjuvant by water
dilution ultrafiltration method.The protein
content of the anti-ultreptococcus inut~zns IgY was
determined by Lowray's method. The titer of the
anti-ultreptococcus rniitans IgY was examined by Passive
JIemagglutination Assay(PIIA). 40 S-D rats have been
established dental caries model were divided into
four groups with random block degisn: the first group
was given 2% the anti-ultreptococcus nzutans IgY in
daily drinks; the second group was given lpptn sodium
fluoride; the third group was given the non-special IgY
extracted by the same method from the eggs laid by the
hens without iniinunity ; the fourth group was given
4
distilled water. After 55 days of treatment, the caries
scores were measured by the keyes's method.
Results: The anti-ultreptococcus fihlltans IgY was produced in eggs
laid by hens after immunity. The protein content of
primary extract of anti-ultreptococcus nzutans IgY
extracted from eggs yolk by water
dilution ultrafiltration method was 100mg/ml. The
titer of this TgY was 1:512. The keyes' caries score of
the first group was significant lower than the one of
the third group and the fourth group(p<0.01) . The
scores of the second group was significant lower than
it of the third group and the fourth group (p<0.01) too.
There wasn't significant difference between the scores
of the first group and the second group.
Conclusion:The primary extract of anti-ultrpotococcus znutans
JgY extracted by the method of this study could
effective prctected rats against dental caries.The
effect was simi larly with sodium fiuorid.
引文
1． 刘天佳主编，口腔疾病的微生物学基础，北京．人民卫生出版社，1999：37-43
2． 樊明文，张平，边专，免疫防龋的研究新进展．中华口腔医学杂志，1999，34(2)：69-72
3. Van Regnmortel MH. Hens could be the antibody factories of the future. World Poultuy,. 1994,10(1/2):67-69
4. Polson A, Von Wechmar MB, Van Regenmortel MH, et al. Isolation of viral IgY antibodies from yolks of immunized hens. Immunol Commun, 1980, 9(5):475-493
5. Hamada S, Horikoshi T, Minami T, et al. Protection of rats against dental caries by immunization with hen-egg-yolk antibody(IgY). Infect Immun, 1991,59(11): 4161-4167
6. Otake. S, Nishihara Y, Makimura M, et al. Passive immunization against dental caries in rats by use of hen egg yolk antibodies specific for cell-associated glucosyltransferase of streptococcus mutans. J Dent Res, 1991, 70(3): 162-166
7. Hatta H, Tsuda K, Ozeki M, et al. Passive immunization against dental plaque formation in humans: effect of a mouth rinse containing egg yolk antibody(IgY). Caries Res,1997, 31(4): 268-274
8． 姜崴，赵小琳，宋昌玲，等．防龋卵黄抗体的研制．中国生物制品学杂志，1999，12(1)：23-25
9. FAN Ming-wen, JIANG Qian-zhou, BIAN zhuan. Isolution of egg IgY hens immunized With S.mutans B29-33 and S.Sobrinus 6715.口腔医学纵横杂志，2000，16(3)：193-194
10．肖晓蓉．口腔微生物学及实用技术．北京：北医大协和医大出版社，1993：91-179
11．李富明，罗宗莲，周学东，等．变形链球菌表面蛋白P_1、主动免疫防龋动物实验效果初步观察。华西口腔医学杂志，1997，15(1)：48-50
12. Keyes PH. Dental Caries in the molar feeth of rats Ⅱ a methods for diagnosting and scoring several types of lesions simultaneously. J Dent Res, 1958,37:1088-1099
13． 樊明文主编。牙体牙髓病学．北京：人民卫生出版社，2000：43
14． 岳松龄主编．现代龋病学．北京：北医大协和医大联合出版社，1993：338
15. Lehner T, Cnddwell J, Smith R, et al. Local passive Immunization by Monoclonal Antibodies against streptococcus Antigen Ⅰ/Ⅱ in the prevention of Dental Caries. Infect Immum, 1985, 50(30):796-799
16. michalek SM, Gregory RL,Harmon C, et al. Protection of gnotobiotic rats against dental caries by passive immunization with bovine milk antibodies to streptococcus mutans. Infect Immun, 1987,52:2341-2347
17. Filler SJ, Gregory RL, Michalek SM ,et al. Effect of immune bovine milk on streptococcus mutans in human dental plaque. Archs oral Biol, 1991,36:41-47
18. Yu H, Nakano Y, Yamashita Y, et al. Effect of Antibodies against cell surface protein Antigen PAC-Glucosyltransferase Fusion proteins on Glucan syntbesis and cell Adhesion of streptococeus mutans. Infect Immun, 1997, 65(6):2292-2298
19. Ma JKC ,Hunjan M, Smith R, et al. An Investigation into the Mechanism of protection by Local Passive Immunization with Monoclonal Antibodies against streptococcus mutans. Infect Immun.,1990,58(10):3407-3414
20. Van Raamsdonk M, de soet JJ, Bosch-Tijhof CJ, et al. Effect of antibodies on chemiluminescene and on rilling of streptococcus Sobrinus by polymorphonuclear leukocyes. Oral Microbiol Immunol, 1996,11:254-258
21. Van Raamsdonk M. de soet JJ. de Graff J. et al. Efect of Monoclonal Antibodies on the colonization of Rats by streptococcus sobrinus Caries Res. 1993: 27:31-37
22. de soet JJ, Van Loveren, Lammens AJ ,et al. Differences in Cariogenicity between fresh isolates of streptococcus Sobrinus and Streptococcus mutans. Caries Res.1991: 25:116-122
23． 黄定明(综述)．变形链球菌和远缘链球菌致龋性的研究近况．上海口腔医学，1996，5(2)：98-101
24. Ma JKC, Smith R ,Lehner T, et al. Use of monoclonal antibodies in local passive immunization to prevent colonization of human teeth by streptococcus inutans. Infect Immun, 1987, 55:1274-1278
25． 张平，樊明文，边专，等．单克隆抗体对茸毛链球菌粘附及龋病发生的影响．中华口腔医学杂志，1999，34(2)：109-111
26． 梁景平，刘正，朱敏，等．抗Ⅰ型菌毛单克隆抗体粘附作用机制的研究．现代口腔医学杂志，2000，14(1)：1-3
27． 杨健．抗变形链球菌鸡卵黄抗体对两种变形链球菌粘附的对两种变形链球菌的影响．硕士研究生毕业论文。南昌：江西医学院，2000：
28. Van Raainsdnk M, Van der Mei HC, de JJ, et al. Effect of polyclonal and monoclonal antibodies on surface properities of streptococcus sobrinus. Infect Immun, 1995, 63:1689-1702
29． 文冰．抗变形链球菌鸡卵黄抗体对变形链球菌产酸及葡糖基转移酶的影响．硕士研究生毕业论文．南昌：江西医学院，2000：
30. OHO T, Shimazaki Y, Mitoma M,et al. Bovine milk antibodies against cell Surface protein antigen PAC-glucosyctransferase fusion protein suppress cell adhesion and alter glucan synthesis of streptococcus mutans.J Nutr., 1999 ,129(10):1836-41
31. Smith DJ, King WF, Godiska R, et al. Passive Transfer of Immunoglobulin Y Antibody to streptococcus mutans Glucan Binding protein B Confer protection against Experim Dental caries. Infect Immun ,2001,69(5):3135-43
32. Gregory RL, Suzane m. Function of anti-streptococcus mutans antibodies:anti-ribosomal antibodies inhibit acid production, growth and glucose phosphotransferase activity.Infect Immun , 1984,45(1):286
33．文立亚．单克隆抗体对变链菌产酸的影响．牙体牙髓牙周病学志，1996，6(4)：196-197
34．史久成，史俊南主编．口腔内科临床新进展．西安：世界图书出版社，2000：57
35．盛江筠，刘正．变形链球菌耐氟菌株脱矿能力的体外研究．牙体牙髓牙周病学 杂志，2000，10(1)：13-15
36．唐镇(综述)．鸡卵黄抗体及其在医学中的运用．中国临床医药研究杂志，2000，12(9)：1352-1353
37．江千舟(综述)．鸡卵黄抗体及其在医学中的运用．国外医学口腔分册，2000，27(3)：179-180
2． 樊明文，张平，边专，免疫防龋的研究新进展．中华口腔医学杂志，1999，34(2)：69-72
3. Van Regnmortel MH. Hens could be the antibody factories of the future. World Poultuy,. 1994,10(1/2):67-69
4. Polson A, Von Wechmar MB, Van Regenmortel MH, et al. Isolation of viral IgY antibodies from yolks of immunized hens. Immunol Commun, 1980, 9(5):475-493
5. Hamada S, Horikoshi T, Minami T, et al. Protection of rats against dental caries by immunization with hen-egg-yolk antibody(IgY). Infect Immun, 1991,59(11): 4161-4167
6. Otake. S, Nishihara Y, Makimura M, et al. Passive immunization against dental caries in rats by use of hen egg yolk antibodies specific for cell-associated glucosyltransferase of streptococcus mutans. J Dent Res, 1991, 70(3): 162-166
7. Hatta H, Tsuda K, Ozeki M, et al. Passive immunization against dental plaque formation in humans: effect of a mouth rinse containing egg yolk antibody(IgY). Caries Res,1997, 31(4): 268-274
8． 姜崴，赵小琳，宋昌玲，等．防龋卵黄抗体的研制．中国生物制品学杂志，1999，12(1)：23-25
9. FAN Ming-wen, JIANG Qian-zhou, BIAN zhuan. Isolution of egg IgY hens immunized With S.mutans B29-33 and S.Sobrinus 6715.口腔医学纵横杂志，2000，16(3)：193-194
10．肖晓蓉．口腔微生物学及实用技术．北京：北医大协和医大出版社，1993：91-179
11．李富明，罗宗莲，周学东，等．变形链球菌表面蛋白P_1、主动免疫防龋动物实验效果初步观察。华西口腔医学杂志，1997，15(1)：48-50
12. Keyes PH. Dental Caries in the molar feeth of rats Ⅱ a methods for diagnosting and scoring several types of lesions simultaneously. J Dent Res, 1958,37:1088-1099
13． 樊明文主编。牙体牙髓病学．北京：人民卫生出版社，2000：43
14． 岳松龄主编．现代龋病学．北京：北医大协和医大联合出版社，1993：338
15. Lehner T, Cnddwell J, Smith R, et al. Local passive Immunization by Monoclonal Antibodies against streptococcus Antigen Ⅰ/Ⅱ in the prevention of Dental Caries. Infect Immum, 1985, 50(30):796-799
16. michalek SM, Gregory RL,Harmon C, et al. Protection of gnotobiotic rats against dental caries by passive immunization with bovine milk antibodies to streptococcus mutans. Infect Immun, 1987,52:2341-2347
17. Filler SJ, Gregory RL, Michalek SM ,et al. Effect of immune bovine milk on streptococcus mutans in human dental plaque. Archs oral Biol, 1991,36:41-47
18. Yu H, Nakano Y, Yamashita Y, et al. Effect of Antibodies against cell surface protein Antigen PAC-Glucosyltransferase Fusion proteins on Glucan syntbesis and cell Adhesion of streptococeus mutans. Infect Immun, 1997, 65(6):2292-2298
19. Ma JKC ,Hunjan M, Smith R, et al. An Investigation into the Mechanism of protection by Local Passive Immunization with Monoclonal Antibodies against streptococcus mutans. Infect Immun.,1990,58(10):3407-3414
20. Van Raamsdonk M, de soet JJ, Bosch-Tijhof CJ, et al. Effect of antibodies on chemiluminescene and on rilling of streptococcus Sobrinus by polymorphonuclear leukocyes. Oral Microbiol Immunol, 1996,11:254-258
21. Van Raamsdonk M. de soet JJ. de Graff J. et al. Efect of Monoclonal Antibodies on the colonization of Rats by streptococcus sobrinus Caries Res. 1993: 27:31-37
22. de soet JJ, Van Loveren, Lammens AJ ,et al. Differences in Cariogenicity between fresh isolates of streptococcus Sobrinus and Streptococcus mutans. Caries Res.1991: 25:116-122
23． 黄定明(综述)．变形链球菌和远缘链球菌致龋性的研究近况．上海口腔医学，1996，5(2)：98-101
24. Ma JKC, Smith R ,Lehner T, et al. Use of monoclonal antibodies in local passive immunization to prevent colonization of human teeth by streptococcus inutans. Infect Immun, 1987, 55:1274-1278
25． 张平，樊明文，边专，等．单克隆抗体对茸毛链球菌粘附及龋病发生的影响．中华口腔医学杂志，1999，34(2)：109-111
26． 梁景平，刘正，朱敏，等．抗Ⅰ型菌毛单克隆抗体粘附作用机制的研究．现代口腔医学杂志，2000，14(1)：1-3
27． 杨健．抗变形链球菌鸡卵黄抗体对两种变形链球菌粘附的对两种变形链球菌的影响．硕士研究生毕业论文。南昌：江西医学院，2000：
28. Van Raainsdnk M, Van der Mei HC, de JJ, et al. Effect of polyclonal and monoclonal antibodies on surface properities of streptococcus sobrinus. Infect Immun, 1995, 63:1689-1702
29． 文冰．抗变形链球菌鸡卵黄抗体对变形链球菌产酸及葡糖基转移酶的影响．硕士研究生毕业论文．南昌：江西医学院，2000：
30. OHO T, Shimazaki Y, Mitoma M,et al. Bovine milk antibodies against cell Surface protein antigen PAC-glucosyctransferase fusion protein suppress cell adhesion and alter glucan synthesis of streptococcus mutans.J Nutr., 1999 ,129(10):1836-41
31. Smith DJ, King WF, Godiska R, et al. Passive Transfer of Immunoglobulin Y Antibody to streptococcus mutans Glucan Binding protein B Confer protection against Experim Dental caries. Infect Immun ,2001,69(5):3135-43
32. Gregory RL, Suzane m. Function of anti-streptococcus mutans antibodies:anti-ribosomal antibodies inhibit acid production, growth and glucose phosphotransferase activity.Infect Immun , 1984,45(1):286
33．文立亚．单克隆抗体对变链菌产酸的影响．牙体牙髓牙周病学志，1996，6(4)：196-197
34．史久成，史俊南主编．口腔内科临床新进展．西安：世界图书出版社，2000：57
35．盛江筠，刘正．变形链球菌耐氟菌株脱矿能力的体外研究．牙体牙髓牙周病学 杂志，2000，10(1)：13-15
36．唐镇(综述)．鸡卵黄抗体及其在医学中的运用．中国临床医药研究杂志，2000，12(9)：1352-1353
37．江千舟(综述)．鸡卵黄抗体及其在医学中的运用．国外医学口腔分册，2000，27(3)：179-180