Methods and compositions for detecting human tumors

Abstract

Methods and compositions for detecting the presence of tumors are provided, where a physiological sample is assayed for the expression product of a c-onc gene as diagnostic for the presence of the tumor. The method finds use in both pre-and postoperative situations with a host suspected of having transformed malignant cells.

Description

RELATED APPLICATIONS

This application is a continuation-in-part application of copending application Ser. No. 439,252, filed Nov. 4, 1982, demonstrate the ability of such nucleic acids to transform vertebrates to malignancy. One may then use these nucleic acids to deduce peptide composition and screen malignant cells for transcripts or peptides, by hybridization in the former case and with appropriate receptors in the latter case, employing any of a wide variety of diagnostic assays. Antibodies can be produced to the peptides, which antibodies may be labeled and may then be used for diagnosing the presence of a peptide diagnostic of malignancy. The oncogernic proteins are found to be available for binding to antibodies as surface membrane proteins. The antibodies may serve as diagnostic reagents for determining the presence of malignancy and determining the location of malignant cells. The antibodies may also serve in treating tumors in vivo by using radionuclides, toxins, in combination with the host complement system or opsonins, or other antibody dependent lytic system or the like. The antibodies find use in pre- and postoperative systems, in the later determining whether complete removal has occurred, whether metastases exist. The antibodies can be used postoperatively to destroy any remnants of the tumor which may not have been excised.

Although the foregoing invention has been described in some detail by way of illustration and example for purposes of clarity of understanding, it will be obvious that certain changes and modifications may be practiced within the scope of the appended claims.

Claims

1. A method for evaluating the probability of cellular malignancy in a human host, said method comprising:

bringing into close associate (1) a probe specific for a cellular product, said cellular product being mRNA or its expression product, where said mRNA is complementary to a dna sequence of a retrovirus capable of transforming a normal cell to malignancy and said probe is a nucleic acid sequence capable of duplexing with said mRNA or antibody capable of binding to said expression product, and (2) a source from said human host suspected of containing cellular product; and

determining the level of binding of said probe to said cellular product, wherein an elevated level is indicative of the presence of cellular malignancy.

2. A method according to claim 1, wherein said source is cells from said human host.

3. A method according to claim 1, wherein said source is a physiological fluid from said human host.

4. A method according to claim 1, wherein said dna sequence is selected from the group consisting of the oncogenes src, fps, yes, fos, myc, erb, myb, rel, mos, bas, abl, ras, fes, fms, and sis.

5. A method according to any of claims 1, 2, 3, or 4, wherein said probe is an antibody.

6. A method according to claim 5, wherein said antibody is labeled with a label capable of providing a detectible signal.

7. A method according to claims 1, 2, 3, or 4, wherein said probe is a polynucleotide of at least 14 bases complementary to said mRNA.

8. A method for evaluating the probability of leukemia in a human host, said method comprising:

combining antibodies specific diagnostic for the presence of for the oncogene myb and blood cells from a human host suspected of having leukemia; and

determining the level of binding of said antibodies to said host blood cells as diagnostic of a leukemic host.

9. A method according to claim 8, wherein said antibodies are produced in response to an oligopeptide mimicking a portion of the conformation of the myb protein.

10. A method for substantially eliminating human malignant cells from a combination of human malignant and normal cells, which comprises:

combining under cytotoxic conditions said combination of cells with an antibody specific for an expression product of a dna sequence present in a retrovirus genome or substantially complementary to said dna sequence, which sequence is expressed in said malignant cells as a surface protein; and

isolating normal cells, substantially free of malignant cells.

11. A method according to claim 10, wherein said separation occurs in the presence of complement as said cytotoxic condition.

12. A method according to claim 10, wherein said antibodies are labeled with a radionuclide as said cytotoxic condition.

13. A method according to claims 10, 11, or 12, wherein said dna sequence is the myb gene.

14. A method for treating a human host suspected of having malignant cells, which comprises:

administering to said human host under cytotoxic conditions antibodies to the expression product of a gene, which gene is part of a retrovirus genome capable of inducing malignancy in a normal cell or which gene is substantially complementary to said gene of said retrovirus genome.

15. A method according to claim 14, wherein said cytotoxic condition is the presence of complement.

16. antibodies specific for the expression product of the human oncogenes c-myc, c-fos, c-ras^Ha, c-ras^Ki, c-fes, c-myb, and c-src.

17. antibodies according to claim 16, labeled with a label capable of providing a detectible signal.

18. antibodies according to claim 16, labeled with a

cytotoxic agent.19. An antigenic oligopeptide selected from the class consisting of:

(a) met-ala-phe-ala-his-asn-pro-pro-ala-gly-pro-leu-pro-gly-ala

(b) pro-phe-his-lys-asp-gln-thr-phe-thr-glu-tyr-arg-lsy-met-his-gly-gly-ala-va

(c) pro-phe-his-lys-asp-gln-thr-phe-thr-glu-tyr-arg-lys-met

(d) asp-asn-thr-arg-thr-ser-gly-asp-asn-ala-pro-val-ser-cys-leu-gly-glu

(e) arg-leu-ileu-gly-asp-asn-glu-tyr-thr-ala-arg-gln-gly-ala-lys-phe-pro

(f) trp-arg-arg-asp-pro-glu-glu-arg-pro-thr

(g) arg-leu-lys-lys-ileu-ser-lys-glu-glu-lys-thr-pro-gly-cys-val-lys-ileu-lys- lys

(h) asp-leu-pro-ser-arg-thr-val-asp-thr-lys-gln-ala-gln-glu-leu-ala-arg

(i) met-thr-glu-tyr-lys-leu-val-val-val-gly-ala-ser-gly-val-gly-lys-ser-ala

(j) glu-asp-ileu-his-gln-try-arg-glu-gln-ileu-lys-arg-val-lys-asp-ser-asp-asp

(k) val-arg-glu-ileu-arg-gln-his-lys-leu-arg-lys-leu-asn-pro-pro-asp-glu-ser-g ly-pro

(l) met-thr-gly-tyr-lys-leu-val-val-val-gly-ala-gly-gly-val-gly-lys-ser-ala

(m) val-asp-glu-tyr-asp-pro-thr-ileu-glu-asp-ser-tyr-arg-lys-gln-val

(n) arg-his-ser-thr-ser-ser-ser-glu-gln-glu-arg-glu-gly-gly-arg

(o) asn-gln-gln-thr-arg-glu-phe-val-glu-lys-gly-gly-arg

(p) pro-glu-val-gln-lys-pro-leu-his-glu-gln

(q) ala-ser-pro-tyr-pro-asn-leu-ser-asn-gln-gln-thr-arg

(r) arg-leu-ileu-ala-glu-lys-glu-gln-leu-arg-arg-arg-arg-glu-gln

(s) asn-asn-glu-lys-ala-pro-lys-val-val. 20. antibodies raised to an antigenic polypeptide selected from the class consisting of:

(a) met-ala-phe-ala-his-asn-pro-pro-ala-gly-pro-leu-pro-gly-ala;

(b) pro-phe-his-lys-asp-gln-thr-phe-thr-glu-tyr-arg-lys-met-his-gly-gly-ala-va l;

(c) pro-phe-his-lys-asp-gln-thr-phe-thr-glu-tyr-arg-lys-met;

(d) asp-asn-thr-arg-thr-ser-gly-asp-asn-ala-pro-val-ser-cys-leu-gly-glu;

(e) arg-leu-ileu-glu-asp-asn-glu-tyr-thr-ala-arg-gln-gly-ala-lys-phe-pro;

(f) trp-arg-arg-asp-pro-glu-glu-arg-pro-thr;

(g) arg-leu-lys-lys-ileu-ser-lys-glu-glu-lys-thr-pro-gly-cys-val-lys-ileu-lys- lys;

(h) asp-leu-pro-ser-arg-thr-val-asp-thr-lys-gln-ala-gln-glu-leu-ala-arg;

(i) met-thr-glu-try-lys-leu-val-val-val-gly-ala-ser-gly-val-gly-lys-ser-ala;

(j) glu-asp-ileu-his-gln-tyr-arg-glu-gln-ileu-lys-arg-val-lys-asp-ser-asp-asp;

(k) val-arg-glu-ileu-arg-gln-his-lys-leu-arg-lys-leu-asn-pro-pro-asp-glu-ser-g ly-pro;

(l) met-thr-glu-tyr-lys-leu-val-val-gly-ala-gly-gly-val-gly-lys-ser-ala;

(m) val-asp-glu-tyr-asp-pro-thr-ileu-glu-asp-ser-tyr-arg-lys-gln-val;

(n) arg-his-ser-thr-ser-ser-ser-glu-gln-glu-arg-glu-gly-gly-arg;

(o) asn-gln-gln-thr-arg-glu-phe-val-glu-lys-gly-gly-arg;

(p) pro-glu-val-gln-lys-pro-leu-his-glu-gln;

(q) ala-ser-pro-tyr-pro-asn-leu-ser-asn-gln-gln-thr-arg;

(r) arg-leu-ileu-ala-glu-lys-glu-gln-leu-arg-arg-arg-arg-glu-gln; and

(s) asn-asn-glu-lys-ala-pro-lys-val-val. 21. antibodies according to claim

20, labeled with a label capable of providing a detectible signal. 22. antibodies according to claim 20, labeled with a cytotoxic agent.

23. A method for evaluating the probability of cellular malignancy in a human host, said method comprising:

bringing into close association (1) a probe diagnostic for the presence of a cellular product, said cellular product being mRNA or its expression product, where said mRNA is capable of hybridizing to a dna sequence of a retrovirus capable of transforming a normal cell to malignancy and said probe is a nucleic acid sequence capable of duplexing with said mRNA or antibody capable of binding to said expression product, and (2) a source from said human host suspected of containing cellular product; and

determining the level of said binding of said probe to said cellular product, wherein an elevated level is indicative of the presence of

cellular malignancy. 24. An antibody diagnostic for the presence of the expression product of a human oncogene selected from the group consisting of c-myc, c-fos, c-ras^K, c-fes, and c-myb. 25. An antibody according to claim 24, wherein said human oncogene is c-myc. 26. An antibody according to claim 24, wherein said human oncogene is c-fos. 27. An antibody according to claim 24, wherein said human oncogene is c-ras^K. 28. An antibody according to claim 24, wherein said human oncogene is c-fes. 29. An antibody according to claim 24, wherein said human oncogene is c-myb. 30. An antibody according to claim 24, labeled with a label capable of providing a detectable signal. 31. An antibody according to claim 24, labeled with a cytotoxic agent.