Physicochemical studies of the carcinoembryonic antigens of the human digestive system.

A procedure has been described for the purification of the carcinoembryonic antigens (CEA) of the human digestive system. Tumor tissue extraction in 0.6 M perchloric acid followed by paper block electrophoresis and column chromatography on Sephadex G-200 resulted in highly purified CEA preparations as determined by both immunological and physicochemical criteria. The properties and composition of five different purified CEA preparations derived from digestive system cancer metastases were examined. The findings demonstrated a high degree of uniformity amongst these samples. Sedimentation coefficients ranged from 6.9 to 8S. Each sample showed the presence of 14 different amino acid residues and six different carbohydrate constituents (four of which could be quantitated with the amount of material available for analyis). Studies of a purified CEA preparation from a primary hepatoma yielded results which, in some respects, differed from those obtained with the CEA samples of metastatic tumor origin. The implications of these variations were discussed with regard to the probable presence of non-CEA components in the hepatoma preparation. Of primary importance was the observation that the few normal adult digestive system tissues tested failed to show the presence of constituents similar to the CEA. This finding would seem to indicate that, in the adult, the carcinoembryonic antigens of the human digestive system are qualitatively tumor-specific and are not dectectable in comparable normal tissues.

The isolation and characterization of tumor-specific antigens of rodent and human tumors.

Putative tumor-specific transplantation antigens (TSTA) from both a carcinogen-induced rodent tumor (MC-1) and 2 human tumors were purified. The antigens were solubilized from the tumor cell membranes by limited papain digestion in a manner similar to that described for the isolation of normal histocompatibility antigens. The antitumor immune response of the tumor-bearing host was used to monitor the purification of the putative TSTA in both the rodent and human tumor systems. In the case of the rodent tumor, a major step in the purification of the TSTA involved affinity chromatography on Sepharose beads coupled to autologous antitumor antiserum. A comparable procedure was utilized in the purification of the TSTA from human tumors by using affinity chromatography on anti-human beta2-microglobulin antiserum coupled to a solid phase. The data obtained indicate that the TSTA of human tumors contains a beta2-microglobulin chain that is immunochemically identical with, and very similar in size to, that found in normal human histocompatibility antigens. A subunit of similar size was also identified in the carcinogen-induced rodent tumor. These results suggest that the TSTA in both humans and rodents may well be altered histocompatibility antigens.

The detection of alpha 1-fetoprotein in patients with viral hepatitis.

PIP: Alpha fetoprotein (AFP) was detected in sera (351 samples) of 128 patients with viral hepatitis by radioimmunoassay. 77 positive tests for AFP were obtained. These positive results were demonstrated on 1 or more samples taken from 40 (31%) of the 128 patients studied; the highest value obtained was 4400 ng/ml. Hepatitis B antigen (HBAg) was positive in 26/40 (65%) of patients in whom AFP was detected during the disease process. However, 58/88 (66%) who were seronegative for AFP also demonstrated HBAg in their sera. Chi-square analysis revealed no significant difference in occurrence of detectable AFP between HBAg seropositive and seronegative patients. Individuals seropositive for AFP had no statistically different concentration of the protein than patients seropositive or seronegative for HBAg. 24 patients' sera were tested serially over a 2-week period. Both the peak glutamic-pyruvic transaminase (GPT) and peak total bilirubin levels were in a higher range in those 10/24 patients seropositive (P .001) for AFP than in the 14/24 who were seronegative. Appearance of AFP was related to the severity of liver tissue destruction, as reflected by serum GPT. However, peak AFP levels were attained 5-16 days after peak serum GPT appeared in the circulation.^ieng

Physicochemical approach to the purification of human alpha1-fetoprotein from the ascites fluid of a hepatoma-bearing patient.

A method for the purification of human alpha1-fetoprotein from the ascites fluid of a hepatoma-bearing patient is described that is capable of yielding large quantities of pure alpha1-fetoprotein within a relatively short period of time. The technique is based entirely on the physicochemical properties of the alpha1-fetoprotein molecule and uses sequential purification steps: ion-exchange chromatography on DEAE-Sephadex A-50, molecular-sieve chromatography on Sephadex G-200, negative-affinity chromatography on Sepharose-Blue Dextran, positivepaffinity chromatography on concanavalin A-Sepharose and, finally, molecular-sieve chromatography on Sephadex G-100. The efficiency of the entire procedure in its present form is 15% of the alpha1-fetoprotein activity of the starting preparation from ascites fluid. The purity of the final product was shown by polyacrylamide gel electrophoresis, radioimmunoelectrophoresis, and determinations of the NH2-terminal and COOH-terminal amino acid residues of the alphs1-fetoprotein isolated. Amino acid analysis of the final product revealed a composition very similar to those reported for alpha-fetoprotein preparations that have been previously isolated by the use of immunochemical technology.

The isolation of the gamma subunit of fetal hemoglocin (HbF) and its use in a radioimmunoassay for HbF.

A method is described for the purification, from fetal hemoglobin (HbF), of the fetal specific globin chain (gamma chain) in its native state. In the absence of alpha chain (the globin chain common to all adult human hemoglobins) gamma chain, when used as an immunogen, is able to express its unique antigenicity. Here, a specific, high titer antiserum raised against gamma chain has been used to establish a sensitive radioimmunoassay for HbF. This approach may be applicable to the measurement of other normal and abnormal hemoglobins.

Oncofetal antigens. Increasing the specificity of the CEA radioimmunoassay.

The biologic and clinical significance of the oncofetal antigens carcinoembryonic antigen (CEA) and alpha1-fetoprotein (AFP) are discussed. Although the current assays for these molecules are not tumor-specific, measurement of these molecules in the circulation of cancer patients is useful either for tumor diagnosis or for management of the cancer patient in the postoperative or post-chemotherapy state. An approach to increasing the specificity of the CEA radioimmunoassay is described.

New perspectives in allergic asthma.

Major advances have recently been made in both the diagnosis and the treatment of allergic asthma. The radioallergosorbent test (RAST), which measures allergen specificity of lgE antibodies in vitro, offers as good results as direct skin tests for allergy, without the inconvenience, discomfort and risk for the patient of the latter. However, the RAST must be done in a radioisotope laboratory, and standardized extracts of test allergens are lacking. Several new drugs, administered by inhalation and relatively free from serious side effects, are highly effective in treating asthma. Disodium cromoglycate, believed to stabilize tissue mast cells, protects particularly against exercise- or antigen-induced bronchospasm, and often permits reduction or withdrawal of corticosteroid therapy. Some acetonides or esters of glucocorticoids, because of their greatly increased topical anti-inflammatory activity, are effective in doses too small to cause serious systemic side effects. With the most topically effective, beclomethasone dipropionate, systemic corticosteroid therapy can usually be reduced or withdrawn, but recovery of hypothalamic-pituitary-adrenal function may be delayed; the drug regimens should therefore overlap.

Double-blind trial comparing flunisolide and placebo for the treatment of perennial rhinitis.

The effectiveness of a new topical steroid, flunisolide, was evaluated for the treatment of patients with perennial rhinitis. The sixty-nine patients in this 8 week double-blind parallel trial received either 300 microgram/day of flunisolide or placebo. The flunisolide group showed statistically greater improvement than the placebo group in such symptoms as the duration of sneezing, stuffy nose, runny nose and nose blowing. 63% of the flunisolide group compared with 39% of the placebo group felt the test spray had provided substantial or total control of their nasal symptoms (P = 0.0026). When those patients who were skin test-positive (allergic) were considered separately from those who were skin test-negative (non-allergic), differences in responsiveness were noted. While those patients who had negative skin tests received greater relief of their symptoms with flunisolide than with placebo, patients who had positive skin tests showed a greater responsiveness to flunisolide and reported a higher incidence of significant or total control of their symptoms. Careful monitoring of plasma cortisol levels revealed no evidence of adrenal suppression in any of the patients in this trial. This lack of suppression may be due to two factors: (a) flunisolide is rapidly metabolized into a metabolite with a low degree of activity and (b) a very low dose of corticosteroid is administered.

Studies of the linkage relationship of beta-2-microglobulin in man-mouse somatic cell hybrids.

Beta-2-microglobulin (beta2mu) production has been studied in 33 primary man-mouse hybrid clones and in 26 secondary man-mouse hybrid clones. These clones have also been examined for the presence of 15 human enzyme phenotypes. Karyotypic analyses have been carried out on clones. From these studies the following conclusions can be drawn: (1) Gene(s) determining human beta2mu production in humans are apparently syntenic with the MPI gene on chromosome 15. (2) Long-term fibroblast lines may be of limited use in mapping studies as chromosomal rearrangements frequently occur in these lines. (3) The gene(s) determining beta2mu production in humans segregate independently of chromosome 6. If the assignment of the genes determining HL-A alloantigens to chromosome 6 is correct, our results imply that beta2mu and HL-A alloantigens are determined by genes carried on different chromosomes, despite the fact that beta2mu forms an integral part of the HL-A molecule.

Radioimmunoassay for human alpha 1 -fetoprotein.

A method of radioimmunoassay has been developed for the quantitation of alpha(1)-fetoprotein in human serum. The assay requires 20 mul of serum, can be completed in 8 hr, and can reproducibly detect concentrations of 20 ng of alpha(1)-fetoprotein per ml of serum. Hence, the method is about 500-fold more sensitive for the detection of alpha(1)-fetoprotein than the Ouchterlony teohnique currently in general use. The procedure is of potential clinical value as an aid in the diagnosis of cancer and a number of noncancerous hepatic diseases.

The radioimmunoassay of circulating carcinoembryonic antigen of the human digestive system.

A radioimmunoassay has been developed for determining the serum levels of carcinoembryonic antigen of the human digestive system in patients with cancer of the colon and rectum. The assay is simple to perform and has a high degree of reproducibility and specificity. The test detects a concentration of 2.5 ng of carcinoembryonic antigen per ml of serum and this has provided the first demonstration of a circulating tumor-specific antigen in the sera of cancer patients.

Vascular involvement in relapsing polychondritis.

Review of four cases of relapsing polychondritis (RP) seen at one hospital in the 12-year period 1963 to 1974 revealed that one patient had aortic insufficiency with large artery involvement, two others had involvement of medium and large arteries and the fourth may have had mucocutaneous vasculitis. Valvular disease has occurred in 9% of all cases of RP reported in the literature and, if vasculitis beyong the aortic root is included, 25% of cases of RP manifested inflammatory vascular disease. The frequency of pseudotumour of the orbit and cochlear-labyrinthine dysfunction is also high and may be a manifestation of vasculitis.

Carcinoembryonic antigen and alpha 1-fetoprotein in ulcerative colitis and regional enteritis.

Sera were collected from 108 patients with inflammatory bowel disease and assayed for carcinoembryonic antigen (CEA) and alpha(1)-fetoprotein (AFP). Seven (14%) of 51 patients with ulcerative colitis had a positive test for CEA and one of these had associated carcinoma of the colon. Ten (19%) of 52 patients with regional enteritis were also seropositive. The sera of 4 (9%) of 47 patients with ulcerative colitis and 2 (5%) of 41 patients with regional enteritis contained small amounts of AFP. Of two unclassified patients one had a positive CEA and the other a positive AFP. No serum was positive for both CEA and AFP. In addition, multiple samples were available for sequential analysis in eight CEA-positive patients but there was no apparent relationship between seropositivity and disease activity. Continued follow-up is now in progress to determine the significance of detectable fetal antigen levels in inflammatory bowel disease.