Monoclonal antibodies and synthetic tumor-associated glycoconjugates in the study of the expression of Thomsen-Friedenreich-like and Tn-like antigens on human cancers.

Synthetic carbohydrate haptens, which are conjugated to carrier human serum albumin molecules [synthetic tumor-associated glycoconjugates (S-TAGs)], were used to immunize mice for monoclonal antibody (MoAb) production. Two of the S-TAGs were composed of haptens related to the Thomsen-Friedenreich (TF) antigen, and their structures are beta Gal(1----3)-beta GalNAc (TF-beta) and beta Gal(1----3) alpha GalNAc (TF-alpha) (Gal = galactose; GaNAc = N-acetylgalactosamine). The third S-TAG was made up of Tn hapten groups of the structure alpha GalNAc-O-serine. MoAbs specific for TF-alpha and Tn were able to be generated. All MoAbs generated against TF-beta cross-reacted with TF-alpha but not with Tn. None of the TF-alpha-specific MoAbs reacted with human carcinomas, whereas several TF-beta and Tn MoAbs were found to react with most human lung, colon, and breast carcinomas. It is believed that this is the first report of the use of synthetic carbohydrate cancer antigens for the production of anticancer MoAbs.

A novel marker for basal (stem) cells of mammalian stratified squamous epithelia and squamous cell carcinomas.

We have developed a monoclonal antibody (174H.64) which selectively recognizes antigens shared by the basal cells of mammalian stratified squamous epithelium and squamous cell carcinoma (SCC). Histopathological studies of the frozen tissue sections demonstrated selective binding of this antibody to SCCs of human, bovine, canine, feline, and murine origin. Tumors of other histological types did not show reactivity with the antibody. In well-differentiated SCCs the peripheral layer of the tumor showed preferential binding of the antibody, suggesting that the antigens are associated with the proliferative compartment of the tumor. Studies on normal human tissues showed selective binding of the antibody to the basal layer of stratified squamous epithelia, thymic epithelial cells, and myoepithelial cells around breast ducts, while no antibody binding was observed for the suprabasal layers of stratified epithelia, simple epithelia, or tissues of nonepithelial origin. A similar pattern of antibody binding was also observed for bovine and murine skin with staining of the basal layer. The antigens detected by monoclonal antibody 174H.64 were characterized from cytoskeletal protein extracts of normal human keratinocytes as well as human and bovine SCC tissues by using an immunoblotting technique. The antigens detected in normal human keratinocytes consisted of two major protein bands of approximate molecular weights of 48,000-50,000 and 57,000. In bovine SCC tumor the antigen detected was the Mr 48,000-50,000 band and in the human SCC tumor it was the Mr 57,000 band. A murine lung SCC model was developed with a murine SCC cell line KLN-205. The lung tumor obtained was reactive against the antibody and showed selective staining of the peripheral layer of the tumor containing the stem cell population. The antigens described by monoclonal antibody 174H.64 appear to be molecules associated with the stem cell populations of normal stratified epithelium and squamous cell carcinoma.

Renal tubular receptor imaging with iodine-131-labeled peanut lectin: pharmacokinetics and renal clearance mechanism in animals.

Intravenously administered peanut lectin (PNA), iodinated with 131I ([131I]PNA), is rapidly cleared from the plasma by the kidneys in dogs (clearance [total body] = 17.52 +/- 8.74 ml/min). Dynamic gamma camera renal scintigraphy demonstrated renal accumulation and excretion phases of the [131I]PNA renogram in dogs and rabbits (% injection dose-at-peak = 21.8 +/- 3.3% and 19.6 +/- 4.3%, time-to-peak = 44.6 +/- 4.8 min and 37.2 +/- 6.9 min, respectively). Immunoperoxidase staining of kidney sections, following i.v. administered PNA, demonstrated predominant accumulation by the proximal tubules of mice, rabbits, and dogs. The basement membrane was intensely stained at early times p.i. while intracellular and luminal PNA was evident within 1 hr. Urine analysis confirmed the presence of intact [131I]PNA in the bladder contents, while protein degradation products, and a small percentage of the free iodide (less than 5%) were noted within 1 hr p.i. The relative proportion of free iodide increased at later times p.i. (greater than 6 hr). A receptor mediated excretion mechanism is proposed for the clearance of PNA and may be useful for the study of renal tubular function.

Deep scopulariopsosis: a case report and sensitivity studies.

A 36-year-old female was admitted to hospital for debridement of chronically inflamed tendon sheaths and adjacent tissues near the left ankle. Despite antibiotic therapy and initial surgical interventions, the inflammation had progressed slowly over 16 months. Histopathological examination of excised tissues in September 1973 revealed a chronic granulomatous inflammation of tendon sheaths and muscle. Many branched hyphal segments, intercalary swollen cells, and a few conidia-like bodies were seen in sections, and also in KOH- and PAS-stained slides prepared from homogenized tissues. Culture of homogenized tissues yielded pure colonies of Scopulariopsis brevicaulis. Sensitivity tests were initially begun with amphotericin B, potassium iodide, and potassium tartrate (0.05-15 mug/ml of the phytone-yeast extract agar), and no inhibitory effect was observed. Subsequently, amphotericin B, antimony, 5-fluorocytosine (5-FC), griseofulvin, hamycin, and mycostatin were tested (25-300 mug/ml of the phytone-yeast extract agar). Of these chemicals, griseofulvin and hamycin proved to be most effective. Antimony and 5-FC were ineffective, and mycostatin produced a negligible effect on growth. The four strains of Lysobacter antibioticus, the producer of myxin antibiotic, strongly inhibited the growth of the fungus.

Vitamin B12 (cobalamin) and folate blood levels in geriatric reference group as measured by two kits.

For serum vitamin B12 levels there was little apparent difference between a geriatric healthy reference group and a hospitalized group for the total population studied; however, the hospitalized males did have an increased prevalence of values less than normal range. The frequency distribution for both sexes of the geriatric reference group gave lower range limits than manufacturer's normal range. (68-632 vs 133-708 pmol/L for Becton Dickinson, and 125-609 vs 179-930 pmol/L for Bio-Rad, using 95% non-parametric limits). For folate there was an increased incidence in values of less than normal in the hospitalized group versus the geriatric reference group, but there was no difference in the ranges calculated for the latter compared to either manufacturer's normal range derived from a younger population. Comparison of results by two manufacturers' kit methods confirmed Bio-Rad's claim to increased low-end sensitivity of standard curve in range of clinical interest.

Adjuvant BCG immunotherapy for malignant melanoma.

A total of 199 patients with stage I malignant melanoma at Clark's level 3 to 5 of invasion were entered into a prospectively controlled randomized clinical trial that attempted to assess the value of local and systemic immunotherapy with BCG (bacille Calmette-Guérin) after surgery. The patients were randomly assigned, with stratification by Clark's level, to receive either routine follow-up or immunotherapy with BCG, administered intradermally with a Heaf gun around the site of wide excision and then given orally for 2 years. Intradermal administration of BCG was repeated after 1 year's oral therapy with BCG. Of the 99 patients in the treatment group 66 had Clark's level 3, 28 had level 4, and 5 had level 5 invasion. Of the 100 patients in the control group, 61 had level 3, 36 had level 4, and 3 had level 5 invasion. Other prognostic factors, such as sex, depth of invasion, histologic features, site of disease and type of surgery, were evenly distributed. There were 57 recurrences of the melanoma, 24 in the treatment group and 33 in the control group. However, this trend was not statistically significant (p = 0.194). The suggestion that BCG may reduce the likelihood of local/regional recurrence has not been confirmed with longer follow-up. There were 13 such recurrences in the BCG group, compared with 21 in the control group; the proportions of patients in each group who had such a recurrence were not significantly different. Of the 199 patients 41 died, 24 in the control group and 17 in the treatment group; again, this difference was not significant. While there may be minor activity in selected patients, there appeared to be no benefit from this form of adjuvant BCG therapy in patients with malignant melanoma.

Monoclonal antibody against a cryptic carbohydrate antigen of murine and human lymphocytes. I. Antigen expression in non-cryptic or unsubstituted form on certain murine lymphomas, on a spontaneous murine mammary carcinoma, and on several human adenocarcinomas.

This paper describes an IgM monoclonal antibody (49H.8) which was produced following immunization of BALB/c mice with human neuraminidase-treated erythrocytes (NE-RBC). 49H.8 reacts with NE-RBC, neuraminidase-treated T lymphocytes (NE-T) and NE B lymphocytes of both human and murine origin. Little or no reactivity with untreated T or B cells could be detected. Thus the 49H.8 antigen is "cryptic" in most normal lymphocytes of both humans and mice. In contrast, the 49H.8 antigen was detected in non-cryptic or unsubstituted form on many non-treated murine lymphomas of both B- and T-cell origin, on the spontaneous murine mammary carcinoma, TA3-HA and on several human adenocarcinomas. The 49H.8 antigen appears to be related to the previously described 49H.24 antigen as shown by sugar inhibition experiments. 49H.24 reacts most strongly with the synthetic disaccharide (betaGa1 (I leads to 3)alpha Ga1NAc) but not at all with beta Ga1(I leads to 3)beta Ga1Nac. 49H.24 does not react with any of the murine or human tumors tested. 49H.8 reacts with both the alpha and beta forms of the disaccharide but reacts most strongly with phenyl-beta-galactoside-containing compounds. In contrast, phenyl-alpha-galactoside-containing compounds produced no reaction. The natural determinant detected by this antibody was not determined but various possibilities are considered. 49H.8 was used to detect antigen apparently shed from growing TA3-Ha cells into the serum and ascites of tumor-bearing mice. These observations suggest that the 49H.8 monoclonal antibody will be valuable as a specific reagent for a common tumor-associated antigen shared by certain murine and human tumors, and as a means of assaying shed tumor antigen in circulation as in the TA3-Ha mammary adenocarcinoma model.