Polypeptides and immunizing compositions containing gram positive polypeptides and methods of use
The present invention provides isolated polypeptides isolatable from a Staphylococcus spp. Also provided by the present invention are compositions that include one or more of the polypeptides, and methods for making and methods for using the polypeptides.
1. A method comprising:
administering an effective amount of a composition to a subject at risk of developing lesions caused by infection by Staphylococcus aureus , the composition comprising:
at least five isolated polypeptides having molecular weights of 88 kDa, 55 kDa, 38 kDa, 37 kDa, 36 kDa, 35 kDa, or 33 kDa, wherein molecular weight is determined by electrophoresis on a sodium dodecyl sulfate-polyacrylamide gel;
wherein the isolated polypeptide having a molecular weight of 88 kDa comprises an amino acid sequence having at least 95% identity to the amino acid sequence of a reference polypeptide, wherein the reference polypeptide has a molecular weight of 88 kDa as determined by sodium dodecyl sulfate-polyacrylamide gel electrophoresis, comprises the amino acid sequences depicted in SEQ ID NO:1, SEQ ID NO:2, SEQ ID NO:3, SEQ ID NO:4, SEQ ID NO:5, SEQ ID NO:6, SEQ ID NO:7, SEQ ID NO:9, SEQ ID NO:10, SEQ ID NO:11, SEQ ID NO:12, SEQ ID NO:13, SEQ ID NO:14, SEQ ID NO:15, SEQ ID NO:16, SEQ ID NO:17, SEQ ID NO:19, SEQ ID NO:20, SEQ ID NO:21, SEQ ID NO:22, SEQ ID NO:24, SEQ ID NO:25, SEQ ID NO:26, SEQ ID NO:27, SEQ ID NO:28, SEQ ID NO:29, SEQ ID NO:30, SEQ ID NO:31, SEQ ID NO:32, and SEQ ID NO:33, and is expressed by Staphylococcus aureus ATCC strain 19636 at a greater level when grown in medium comprising 1600 μM 2,2-dipyridyl compared to when grown in the medium without the 2,2-dipyridyl;
wherein the isolated polypeptide having a molecular weight of 55 kDa comprises an amino acid sequence having at least 95% identity to the amino acid sequence of a reference polypeptide, wherein the reference polypeptide has a molecular weight of 55 kDa as determined by sodium dodecyl sulfate-polyacrylamide gel electrophoresis, comprises the amino acid sequences depicted in SEQ ID NO:35, SEQ ID NO:37, SEQ ID NO:39, SEQ ID NO:40, SEQ ID NO:41, SEQ ID NO:42, SEQ ID NO:43, SEQ ID NO:44, SEQ ID NO:45, SEQ ID NO:46, SEQ ID NO:49, SEQ ID NO:51, SEQ ID NO:52, SEQ ID NO:53, SEQ ID NO:54, SEQ ID NO:57, SEQ ID NO:58, and SEQ ID NO:59, and is expressed by Staphylococcus aureus ATCC strain 19636 at a greater level when grown in medium comprising 1600 μM 2,2-dipyridyl compared to when grown in the medium without the 2,2-dipyridyl;
wherein the isolated polypeptide having a molecular weight of 38 kDa comprises an amino acid sequence having at least 95% identity to the amino acid sequence of a reference polypeptide, wherein the reference polypeptide has a molecular weight of 38 kDa as determined by sodium dodecyl sulfate-polyacrylamide gel electrophoresis, comprises the amino acid sequences depicted in SEQ ID NO:64, SEQ ID NO:65, SEQ ID NO:66, SEQ ID NO:68, SEQ ID NO:69, SEQ ID NO:70, SEQ ID NO:72, SEQ ID NO:75, SEQ ID NO:76, and SEQ ID NO:77, and is expressed by Staphylococcus aureus ATCC strain 19636 at a greater level when grown in medium comprising 1600 μM 2,2-dipyridyl compared to when grown in the medium without the 2,2-dipyridyl;
wherein the isolated polypeptide having a molecular weight of 37 kDa comprises an amino acid sequence having at least 95% identity to the amino acid sequence of a reference polypeptide, wherein the reference polypeptide has a molecular weight of 37 kDa as determined by sodium dodecyl sulfate-polyacrylamide gel electrophoresis, comprises the amino acid sequences depicted in SEQ ID NO:80, SEQ ID NO:82, SEQ ID NO:84, SEQ ID NO:85, SEQ ID NO:86, SEQ ID NO:87, SEQ ID NO:89, SEQ ID NO:90, SEQ ID NO:91, SEQ ID NO:94, SEQ ID NO:95, SEQ ID NO:96, SEQ ID NO:97, SEQ ID NO:99, SEQ ID NO:100, and SEQ ID NO:101, and is expressed by Staphylococcus aureus ATCC strain 19636 at a greater level when grown in medium comprising 1600 μM 2,2-dipyridyl compared to when grown in the medium without the 2,2-dipyridyl;
wherein the isolated polypeptide having a molecular weight of 36 kDa comprises an amino acid sequence having at least 95% identity to the amino acid sequence of a reference polypeptide, wherein the reference polypeptide has a molecular weight of 36 kDa as determined by sodium dodecyl sulfate-polyacrylamide gel electrophoresis, comprises the amino acid sequences depicted in SEQ ID NO:103, SEQ ID NO:104, SEQ ID NO:105, SEQ ID NO:106, SEQ ID NO:107, SEQ ID NO:108, SEQ ID NO:109, and SEQ ID NO:110, and is expressed by Staphylococcus aureus ATCC strain 19636 at a greater level when grown in medium comprising 1600 μM 2,2-dipyridyl compared to when grown in the medium without the 2,2-dipyridyl;
wherein the isolated polypeptide having a molecular weight of 35 kDa comprises an amino acid sequence having at least 95% identity to the amino acid sequence of a reference polypeptide, wherein the reference polypeptide has a molecular weight of 35 kDa as determined by sodium dodecyl sulfate-polyacrylamide gel electrophoresis, comprises the amino acid sequences depicted in SEQ ID NO:112, SEQ ID NO:113, SEQ ID NO:114, SEQ ID NO:115, SEQ ID NO:116, SEQ ID NO:117, SEQ ID NO:118, SEQ ID NO:119, SEQ ID NO:120, SEQ ID NO:121, SEQ ID NO:122, SEQ ID NO:123, and SEQ ID NO:124, and is expressed by Staphylococcus aureus ATCC strain 19636 at a greater level when grown in medium comprising 1600 μM 2,2-dipyridyl compared to when grown in the medium without the 2,2-dipyridyl; and
wherein the isolated polypeptide having a molecular weight of 33 kDa comprises an amino acid sequence having at least 95% identity to the amino acid sequence of a reference polypeptide, wherein the reference polypeptide has a molecular weight of 33 kDa as determined by sodium dodecyl sulfate-polyacrylamide gel electrophoresis, comprises the amino acid sequences depicted in SEQ ID NO:126, SEQ ID NO:127, SEQ ID NO:128, SEQ ID NO:129, SEQ ID NO:130, SEQ ID NO:131, SEQ ID NO:132, SEQ ID NO:134, SEQ ID NO:135, SEQ ID NO:136, SEQ ID NO:137, SEQ ID NO:138, SEQ ID NO:140, SEQ ID NO:142, and SEQ ID NO:143, and is expressed by Staphylococcus aureus ATCC strain 19636 at a greater level when grown in medium comprising 1600 μM 2,2-dipyridyl compared to when grown in the medium without the 2,2-dipyridyl.
2. The method of claim 1 wherein the subject is a mammal.
3. The method of claim 2 wherein the mammal is a human.
4. The method of claim 2 wherein the mammal is bovine, ovine, porcine, caprine, cervine, bisontine, or equine.
5. The method of claim 1 wherein the subject is avian.
6. The method of claim 1 wherein the subject is a companion animal.
7. The method of claim 1 wherein the composition is administered prophylactically.
8. The method of claim 1 wherein an effective amount is an amount effective to decrease the average size of lesions exhibited by the subject compared to an untreated control.
9. The method of claim 1 wherein an effective amount is an amount effective to decrease the number of lesions exhibited by the subject compared to an untreated control.
10. The method of claim 1 wherein the lesion comprises a necrotic skin lesion.
11. The method of claim 1 wherein the composition comprises the 88 kDa polypeptide, the 55 kDa polypeptide, the 38 kDa polypeptide, the 37 kDa polypeptide, the 36 kDa polypeptide, the 35 kDa polypeptide, and the 33 kDa polypeptide.
12. A method comprising:
administering an effective amount of a composition to a subject at risk of developing a post-surgical wound infection caused by Staphylococcus aureus , the composition comprising:
at least five isolated polypeptides having molecular weights of 88 kDa, 55 kDa, 38 kDa, 37 kDa, 36 kDa, 35 kDa, or 33 kDa, wherein molecular weight is determined by electrophoresis on a sodium dodecyl sulfate-polyacrylamide gel;
wherein the isolated polypeptide having a molecular weight of 88 kDa comprises an amino acid sequence having at least 95% identity to the amino acid sequence of a reference polypeptide, wherein the reference polypeptide has a molecular weight of 88 kDa as determined by sodium dodecyl sulfate-polyacrylamide gel electrophoresis, comprises the amino acid sequences depicted in SEQ ID NO:1, SEQ ID NO:2, SEQ ID NO:3, SEQ ID NO:4, SEQ ID NO:5, SEQ ID NO:6, SEQ ID NO:7, SEQ ID NO:9, SEQ ID NO:10, SEQ ID NO:11, SEQ ID NO:12, SEQ ID NO:13, SEQ ID NO:14, SEQ ID NO:15, SEQ ID NO:16, SEQ ID NO:17, SEQ ID NO:19, SEQ ID NO:20, SEQ ID NO:21, SEQ ID NO:22, SEQ ID NO:24, SEQ ID NO:25, SEQ ID NO:26, SEQ ID NO:27, SEQ ID NO:28, SEQ ID NO:29, SEQ ID NO:30, SEQ ID NO:31, SEQ ID NO:32, and SEQ ID NO:33, and is expressed by Staphylococcus aureus ATCC strain 19636 at a greater level when grown in medium comprising 1600 μM 2,2-dipyridyl compared to when grown in the medium without the 2,2-dipyridyl;
wherein the isolated polypeptide having a molecular weight of 55 kDa comprises an amino acid sequence having at least 95% identity to the amino acid sequence of a reference polypeptide, wherein the reference polypeptide has a molecular weight of 55 kDa as determined by sodium dodecyl sulfate-polyacrylamide gel electrophoresis, comprises the amino acid sequences depicted in SEQ ID NO:35, SEQ ID NO:37, SEQ ID NO:39, SEQ ID NO:40, SEQ ID NO:41, SEQ ID NO:42, SEQ ID NO:43, SEQ ID NO:44, SEQ ID NO:45, SEQ ID NO:46, SEQ ID NO:49, SEQ ID NO:51, SEQ ID NO:52, SEQ ID NO:53, SEQ ID NO:54, SEQ ID NO:57, SEQ ID NO:58, and SEQ ID NO:59, and is expressed by Staphylococcus aureus ATCC strain 19636 at a greater level when grown in medium comprising 1600 μM 2,2-dipyridyl compared to when grown in the medium without the 2,2-dipyridyl;
wherein the isolated polypeptide having a molecular weight of 38 kDa comprises an amino acid sequence having at least 95% identity to the amino acid sequence of a reference polypeptide, wherein the reference polypeptide has a molecular weight of 38 kDa as determined by sodium dodecyl sulfate-polyacrylamide gel electrophoresis, comprises the amino acid sequences depicted in SEQ ID NO:64, SEQ ID NO:65, SEQ ID NO:66, SEQ ID NO:68, SEQ ID NO:69, SEQ ID NO:70, SEQ ID NO:72, SEQ ID NO:75, SEQ ID NO:76, and SEQ ID NO:77, and is expressed by Staphylococcus aureus ATCC strain 19636 at a greater level when grown in medium comprising 1600 μM 2,2-dipyridyl compared to when grown in the medium without the 2,2-dipyridyl;
wherein the isolated polypeptide having a molecular weight of 37 kDa comprises an amino acid sequence having at least 95% identity to the amino acid sequence of a reference polypeptide, wherein the reference polypeptide has a molecular weight of 37 kDa as determined by sodium dodecyl sulfate-polyacrylamide gel electrophoresis, comprises the amino acid sequences depicted in SEQ ID NO:80, SEQ ID NO:82, SEQ ID NO:84, SEQ ID NO:85, SEQ ID NO:86, SEQ ID NO:87, SEQ ID NO:89, SEQ ID NO:90, SEQ ID NO:91, SEQ ID NO:94, SEQ ID NO:95, SEQ ID NO:96, SEQ ID NO:97, SEQ ID NO:99, SEQ ID NO:100, and SEQ ID NO:101, and is expressed by Staphylococcus aureus ATCC strain 19636 at a greater level when grown in medium comprising 1600 μM 2,2-dipyridyl compared to when grown in the medium without the 2,2-dipyridyl;
wherein the isolated polypeptide having a molecular weight of 36 kDa comprises an amino acid sequence having at least 95% identity to the amino acid sequence of a reference polypeptide, wherein the reference polypeptide has a molecular weight of 36 kDa as determined by sodium dodecyl sulfate-polyacrylamide gel electrophoresis, comprises the amino acid sequences depicted in SEQ ID NO:103, SEQ ID NO:104, SEQ ID NO:105, SEQ ID NO:106, SEQ ID NO:107, SEQ ID NO:108, SEQ ID NO:109, and SEQ ID NO:110, and is expressed by Staphylococcus aureus ATCC strain 19636 at a greater level when grown in medium comprising 1600 μM 2,2-dipyridyl compared to when grown in the medium without the 2,2-dipyridyl;
wherein the isolated polypeptide having a molecular weight of 35 kDa comprises an amino acid sequence having at least 95% identity to the amino acid sequence of a reference polypeptide, wherein the reference polypeptide has a molecular weight of 35 kDa as determined by sodium dodecyl sulfate-polyacrylamide gel electrophoresis, comprises the amino acid sequences depicted in SEQ ID NO:112, SEQ ID NO:113, SEQ ID NO:114, SEQ ID NO:115, SEQ ID NO:116, SEQ ID NO:117, SEQ ID NO:118, SEQ ID NO:119, SEQ ID NO:120, SEQ ID NO:121, SEQ ID NO:122, SEQ ID NO:123, and SEQ ID NO:124, and is expressed by Staphylococcus aureus ATCC strain 19636 at a greater level when grown in medium comprising 1600 μM 2,2-dipyridyl compared to when grown in the medium without the 2,2-dipyridyl; and
wherein the isolated polypeptide having a molecular weight of 33 kDa comprises an amino acid sequence having at least 95% identity to the amino acid sequence of a reference polypeptide, wherein the reference polypeptide has a molecular weight of 33 kDa as determined by sodium dodecyl sulfate-polyacrylamide gel electrophoresis, comprises the amino acid sequences depicted in SEQ ID NO:126, SEQ ID NO:127, SEQ ID NO:128, SEQ ID NO:129, SEQ ID NO:130, SEQ ID NO:131, SEQ ID NO:132, SEQ ID NO:134, SEQ ID NO:135, SEQ ID NO:136, SEQ ID NO:137, SEQ ID NO:138, SEQ ID NO:140, SEQ ID NO:142, and SEQ ID NO:143, and is expressed by Staphylococcus aureus ATCC strain 19636 at a greater level when grown in medium comprising 1600 μM 2,2-dipyridyl compared to when grown in the medium without the 2,2-dipyridyl.
13. The method of claim 12 wherein the subject is a mammal.
14. The method of claim 13 wherein the mammal is a human.
15. The method of claim 13 wherein the mammal is bovine, ovine, porcine, caprine, cervine, bisontine, or equine.
16. The method of claim 12 wherein the subject is avian.
17. The method of claim 12 wherein the subject is a companion animal.
18. The method of claim 12 wherein the composition is administered prophylactically.
19. The method of claim 12 wherein the composition comprises the 88 kDa polypeptide, the 55 kDa polypeptide, the 38 kDa polypeptide, the 37 kDa polypeptide, the 36 kDa polypeptide, the 35 kDa polypeptide, and the 33 kDa polypeptide.