Natural human IgG inhibits the production of tumor necrosis factor-α and interleukin-1α through the Fc portion

The overproduction of cytokines such as the tumor necrosis factor- (TNF-) and interleukin-1 (IL-1) may cause further deterioration in the already critical condition of patients with shock, sepsis, and acute inflammation. The effectiveness of infusion therapy of natural human IgG to such patients is suggested to depend partly upon the inhibition of the productivity of these cytokines. In this study, we investigated the modulation effects of IgG and its fragments on the production of TNF- and IL-1, on human peripheral blood mononuclear cells (PBMC). The production of TNF- and IL-1 was found to be dose-dependently inhibited by IgG when stimulated by lipopolysaccharide (LPS), phytohemagglutinin (PHA), concanavalin A (Con A), and interleukin-2 (IL-2). However, no inhibition was seen when stimulated by phorbormyristate acetate (PMA). The F(ab)₂ fragment showed enhancing effects on cytokine production by LPS, while the Fc fragment showed not as much inhibitory effect as whole intact IgG. IgG showed no direct cytotoxic effect on PBMC. These data suggest that natural human IgG inhibits TNF- and IL-1 production by PBMC through the Fc portion. The results of this study led us to conclude that whole intact IgG may be the best form of therapeutic delivery.     

Effect of tumor necrosis factor-α and interferon-γ on the growth of human prostate cancer cell lines

Human recombinant tumor necrosis factor- (rTNF-, 10^-12–10^-8 M) inhibited the proliferation of androgen-dependent LNCaP cells by 32–56%. In contrast, proliferation of androgen-independent PC-3 and JCA-1 cells was only slightly inhibited, or not inhibited at all, respectively. Human recombinant interferon- (rIFN-, 500 U/ml) decreased proliferation of PC-3 and JCA-1 cells by 35% and 53%, respectively, but had no effect on LNCaP cells. Interestingly, the combination of rIFN- and TNF- had greater antiproliferative effects on JCA-1 cells than treatment with either cytokine alone. However, the antiproliferative effects of this combination were similar to those observed for PC-3 or LNCaP cells treated with rIFN- or TNF- alone, respectively. These data suggest that some forms of androgen-independent prostate cancer may benefit from a combination therapy of IFN- and TNF-, while the use of IFN- alone may be more efficacious in others.     

Growth fraction of human bladder tumors

Summary The growth fraction of bladder tumors was immunohistochemically assessed in situ using anti-DNA polymerase (Pol ) monoclonal antibody. This enzyme is known to be present in the nucleus of the cells in G1, S, and G2 phases. The percentage of labeled cells was expressed as the labeling index (LI). The average LI was 6.0% in normal epithelium and 17.8% in bladder tumors, this difference being significant. The labeled cells were distributed throughout both the basal and surface layers of bladder tumors. However, in some bladder tumors, the distribution of Pol -labeled cells varied from area to area. The higher fraction of labeled cells was found in high grade or invasive tumors. Papillary and nodular bladder tumors showed a greater rate of cell proliferation than papillary tumors. These findings suggest that Pol immunostaining could be a potent tool for easy and quick evaluation of proliferating cells in bladder tumors, there-by providing a supplement to conventional histological findings.     

Potential uses of interferon α2 as adjuvant therapy in cancer

Background: The purpose of this study was to provide an overview of the potential uses of adjuvant interferon (IFN) therapy for resected solid tumors at high risk for postsurgical relapse. Methods: A MEDLINE search (1970–1994) of the English-language literature for original articles, reviews, and abstracts addressing IFN use in the adjuvant setting together with the authors' collective experience formed the basis for this review. Results: The use of adjuvant IFN- has been studied most extensively in conjunction with the treatment of melanoma. Fewer data are available on IFN- use for the treatment of other solid tumors. In melanoma, there is evidence from Intergroup trials (Eastern Cooperative Oncology Group and World Health Organization) that IFN-2a given for 1–3 years prolongs the interval to relapse and may have a survival benefit. Trials of adjuvant IFN, with and without chemotherapy, are ongoing in the treatment of renal cell carcinoma and colorectal adenocarcinoma. Its value in the treatment of osteosarcoma and high-grade astrocytoma is unknown. Conclusions: The use of IFN in the adjuvant setting is an exciting area of medical and surgical oncology and has the potential to prolong the time to relapse and to increase survival of patients with melanoma. Its role in the adjuvant therapy of other solid tumors remains to be defined.     

Hyperthermic Isolated Perfusion With Low-Dose Tumor Necrosis Factor α and Doxorubicin for the Treatment of Limb-Threatening Soft Tissue Sarcomas

Background Tumor necrosis factor (TNF)-–based hyperthermic isolated limb perfusion (HILP) is one of the most active available approaches for locally advanced soft tissue sarcomas (STS) of the limbs. The aim of this study was to investigate the anticancer activity of a novel drug regimen including doxorubicin (DXR) and low-dose TNF-.Methods HILP with low-dose TNF- (1 mg) and DXR (8.5 mg/L of limb volume) was given to 21 patients with limb-threatening STS: 14 had primary and 7 had recurrent STS, most of which were high grade (grade 1, n = 3; grade 2, n = 6; grade 3, n = 12). Resection of the tumor remnant was performed 6 to 8 weeks after HILP. TNF- concentrations in plasma and perfusate were measured throughout perfusion.Results A major tumor response was observed at histology and clinical evaluation in 90% and 62% of patients, respectively. After a median follow-up of 30 months, limb salvage and local disease control were achieved in 71% and 81% of cases, respectively. Fourteen patients had moderate regional toxicity, which was resolved in all cases. One patient had severe limb toxicity, which did not require amputation. Systemic side effects were minimal, and there were no postoperative deaths. The perfusate/plasma area under the curve ratio for TNF- was 56.Conclusions HILP with low-dose TNF- and DXR seems to be an active neoadjuvant drug regimen against limb-threatening STS. This therapeutic approach can achieve high limb-sparing surgery rates with acceptable local and negligible systemic toxicity.     

Receptor function studies in specimens from the proximal human urethra obtained by transurethral resection

Summary During transurethral resection (TUR) for prostatic hyperplasia, specimens were taken from the proximal urethra. Muscle strips thus obtained were mounted in an organ bath and muscle contraction was induced by adding increasing concentrations of noradrenaline (NA), methoxamine (₁-agonist) and clonidine (₂-agonist). NA and methoxamine induced a dose-dependent muscle contraction, but clonidine had no effect. The influence of prazosin (₁-antagonist) and yohimbine (₂-antagonist) on the NA-induced muscle contraction was also evaluated. Both antagonists had an inhibitory effect,which was much more potent with prazosin. The specimens taken during TUR were found to be suitable for in vitro receptor function studies. The -adrenergic receptor function in the proximal human urethra was found to be mainly of the -type.     

Growth factors and kidney development

The formation of the metanephric kidney is dependent upon the timed and sequential expression of a number of polypeptide growth factors. To shed light on the participation of members of the insulin-like growth factor (IGF) and epidermal growth factor/transforming growth factor- (EGF/TGF-) families, we measured the synthesis of IGF-I, IGF-II, EGF and TGF- by developing rat metanephroi in organ culture and determined the effect of anti-growth factor antibodies on growth and development. IGF-I, IGF-II and TGF- were produced by metanephroi and released into culture media. We could detect no EGF. Inclusion of anti-IGF-I, anti-IGF-II, anti-IGF-II receptor or anti-TGF- antibodies in organ cultures inhibited growth and development of metanephroi. Our findings suggest that both members of the IGF family and TGF- are produced within the developing metanephros and promote renal organogenesis.     

High amino acid uptake in a low-grade desmoplastic infantile ganglioglioma in a 14-year-old patient

Amino acid uptake is higher in high-grade than in low-grade gliomas; this is the rationale for using radioactively labelled amino acids for the non-invasive grading of brain neoplasms. We present a 14-year-old boy with a low-grade desmoplastic infantile ganglioglioma (DIG) that exhibited marked contrast enhancement on magnetic resonance imaging (MRI), but no signs of infiltration and only minimal surrounding edema. In this benign neoplasm the relative uptake of the radioactively labelled amino acid I-123--methyl tyrosine (IMT), determined using single-photon emission computed tomography (SPECT), was 3.24; it was considerably higher than that of eleven other pretherapeutic low-grade gliomas where it ranged from 1.06 to 1.94 and also markedly above the average value of 2.37 found in 20 high-grade gliomas. This case report illustrates that results from emission tomography with radioactively labelled amino acids must be interpreted with caution, particularly when rare tumor entities are considered in view of uncommon clinical or radiological findings.     

Cytostatic effect of different strains of Bacillus Calmette-Guérin on human bladder cancer cells in vitro alone and in combination with mitomycin C and Interferon-α

Summary The cytostatic activity of five Bacillus Calmette-Guérin (BCG) strains (Pasteur, Evans, Tice, RIVM and Connaught) on human transitional cell cancer T24 cells was examined. A striking effect was noted even in 2-day cultures, and the effect was more pronounced when the cells were incubated for 5 days with different BCG strains alone. The concentrations needed were about the same as those used in clinical practice (10⁹ colonyforming units of Pasteur strain in 100 ml buffered saline solution). Combination with mitomycin C or interferon--2b potentiated the cytostatic effect. A slight difference in cytostatic activity between different BCG strains was found.     

198. Die Resultate der operativen Behandlung des postthrombotischen Syndroms

Zusammenfassung Zur Beseitigung der tiefen Veneninsuffizienz werden wir die Ersatzklappen-Operation an der V. poplitea an. Nach dieser Operation entsteht eine Klappen-funktion, die der normalen Venenklappen entspricht; these führt zu einer Normalisierung oder mindestens zu einer Besserung der venösen Hämodynamik. Die Spätergebnisse beweisen die günstige Beeinflussung der Hämodynamik; die klinische- Untersuchung, die Phlebographie, die Isotopen-Phlebographie and hauptsächlich die vergleichenden prä- and postoperativen Venendruckmessungen untermauern these Tatsache.     

Prostatitissyndrom

Zusammenfassung Das Prostatitissyndrom ist eine multifaktorielle Erkrankung mit weitgehend unbekannter Ätiologie. Völlig unterschiedliche Behandlungskonzepte kommen deshalb zum Einsatz. Entsprechend der neuen NIH-Klassifikation werden pathogene Erreger nur bei der akuten und chronischen bakteriellen Prostatitis ursächlich nachgewiesen. Eine ausreichend lange Antibiotikatherapie, vornehmlich mit Fluorchinolonen, wird dann empfohlen. Die meisten Patienten leiden an einem chronischen Beckenschmerzsyndrom, das in eine entzündliche und eine nichtentzündliche Form unterteilt wird. Ob bei der entzündlichen Form eine Infektion ursächlich ist, ist unklar. Eine probatorische Antibiotikatherapie ist deshalb in der Wirkung umstritten. Bei nachgewiesener oder angenommener funktioneller subvesikaler Obstruktion werden -Rezeptorenblocker empfohlen. Begleitend sollte eine symptomatische, gegen den Beckenschmerz gerichtete Therapie durchgeführt werden. Es ist wichtig, dass der Patient über die Problematik der Diagnose und die Limitierung des Therapieerfolges aufgeklärt ist.     

Characterization of local inflammatory response in an isolated lung perfusion model

Background: Current phase I trials of isolated lung perfusion for treatment of pulmonary metastases have an arbitrarily determined length of perfusion. Our objective was to examine the temporal course of the local and distant inflammatory response as a function of the length of perfusion (ischemia) and subsequent reperfusion in an equivalent animal model. Methods: Sixty male Fischer 344 rats were randomized into four groups (n=15). Each group underwent left isolated lung perfusion with buffered Hespan for 10, 30, 60, or 90 minutes. Subsequently, two subgroups of five animals within each group were allowed to reperfuse for 1 or 3 hours, respectively. Non-perfused right lung was used as control. At each time point, lung specimens were assayed for TNF- by ELISA and histologic sections were examined. Results: There was no significant difference between the left and right lung tissue levels of TNF- at the termination of the ischemic period. However, on reperfusion, the left lung TNF- levels increased significantly above the ischemia baseline in all groups, with a greater magnitude of rise in the groups with 60 and 90 minutes of preceding ischemia (12757 ± 1985 vs. 3524 ± 494 pg/g, and 16914 ± 1657 vs. 6530 ± 1104 pg/g, respectively;p<0.05). There was no significant elevation in tissue levels of TNF- in the right lung. Histologic changes consistent with early pulmonary edema were first detected at 12 hours following onset of reperfusion. Conclusions: Reperfusion following prolonged pulmonary ischemia during isolated lung perfusion results in a significant elevation of local tissue levels of TNF- and may render the perfused lung vulnerable to the adverse effects of the inflammatory cascade.Dr. Burt died October 4, 1997.Address correspondence to the Department of Surgery, Memorial Sloan-Kettering Cancer Center, 1275 York Ave., New York, NY 10021, USA.Presented in part at the 50th Annual Meeting of The Society of Surgical Oncology, March 20–23, 1997, Chicago, IL.     

Urinary unconjugated 5α-androstane-3α, 17β-diol in patients with prostatic tumours

Summary A sensitive and reliable radioimmunoassay (RIA) for urinary unconjugated 5-androstane-3, 17-diol is described. The mean overall recovery of unconjugated 5-androstane-3, 17-diol was found to be 57.4%. The sensitivity of the assay was 79 fmol per assay tube and the intra and inter-assay variations ranged between 7.2% and 11.4%. The mean ± SEM for the concentration of this androgen in the urine of normal men was 339.6±66.8 nmol/24h. The corresponding values for patients with benign prostatic hypertrophy (BHP) and carcinoma of the prostate (Ca) were 297.8±44.7 and 1592.1±622.7 respectively. The mean value for Ca patients was significantly higher than either BPH (p<0.05) or normal subject (p<0.02), suggesting a differential urinary excretion pattern for unconjugated 5-androstane-3, 17-diol between BPH and Ca patients. It is concluded that the combined measurement of this androgen in the plasma and urine provides a more accurate assessment of the profile of this hormone than a single plasma estimation.     

α-Adrenoceptor function before and after chemical sympathectomy in human and feline detrusor muscles

Summary Isolated bladder segments from man and cat were treated with 6-hydroxydopamine (6-OHDA) in vitro. Chemical sympathectomy was evaluated with fluorescence microscopy and found to be very similar to the effect of 6-OHDA administered in vivo to cats. Isometric smooth muscle contractile responses were induced by field stimulation (FS). The amplitude of the responses increased after denervation. The effects of -adrenoceptor agonists and antagonists on the FS-induced contractile responses were compared before and after treatment with 6-OHDA. The reduction in the contractile responses after the addition of noradrenaline to the feline bladder strips was more pronounced after treatment. Phentolamine induced an increase in contractile responses before treatment, an effect not seen afterwards in human bladder strips but which persisted in feline bladder strips. Selective -adrenoceptor agonists did not alter the contractile responses in denervated strips. It is suggested that the function of the -adrenoceptors in the detrusor is to inhibit neuronally mediated contractile responses of smooth muscle.     

Elevated concentrations of the small stress protein HSP27 in rat renal tumors

The expression of two small stress proteins, B crystallin and the 27-kDa heat shock protein (HSP27), was studied quantitatively and immunohistochemically in normal kidney and renal tumors in rats. Levels of B crystallin in renal cell tumors tended to be higher than in normal kidney (P = 0.07), but with a wide range of values, whereas they were significantly lower in mesenchymal tumors (P < 0.0001). In contrast, HSP27 concentrations in both renal cell (mean ± SD: 1790 ± 940 ng/mg protein,n = 15) and mesenchymal (1260 ± 1080 ng/mg protein,n = 10) tumors were significantly higher than the normal kidney value (142 ± 30 ng/mg protein,n = 10,P < 0.0001). A positive correlation was found between B crystallin and HSP27 levels limited to the renal cell tumor case (Pearson's correlation coefficient,r = 0.68,P < 0.01). Immunohistochemistry revealed the loops of Henle to be positive for B crystallin, whereas HSP27 staining was positive in glomerular and interstitial vascular walls and epithelial cells of proximal and distal tubules. Positive immunostaining for B crystallin was demonstrated in six of nine renal cell tumors (67%) studied and for HSP27 in all of the nine cases (100%).     

Role of the E-cadherin/α-catenin complex in modulating cell-cell and cell-matrix adhesive properties of invasive colon carcinoma cells

Background: The clinical behavior of colorectal cancer depends on its ability to invade and metastasize. Metastatic cells must dissociate from other cells and invade through basement membrane and stroma. Cell-cell adhesion in epithelial cells is mediated by the cell surface protein E-cadherin in association with - and -catenin, which link E-cadherin to the cytoskeleton. Decreased cell-cell adhesion and increased motility on laminin have been correlated with more poorly differentiated and aggressive carcinomas. Methods: In this study, the RKO cell line, previously shown by us to lack E-cadherin expression, was transfected with the complementary DNA for E-cadherin. The transfectants were selected for high levels of surface expression by sequential FACS and examined in functional assays. Results: In comparison to control transfectants, the E-cadherin transfectants exhibited a more epithelial-like morphology, a 30% increase in Ca²⁺-dependent cell-cell aggregation, and a markedly reduced motility on the matrix proteins, collagen I and laminin. Conclusions: These data demonstrate that correction of a defect in the cadherin/catenin cell-cell adhesion complex, often found in poorly differentiated and highly invasive tumors, facilitates increased cell-cell adhesion and retards tumor cell migration on basement membrane and stromal proteins.The results of this study were presented at the 47th Annual Cancer Symposium of The Society of Surgical Oncology, Houston, Texas, March 17–20, 1994.     

Transgenic Mice Reveal Roles for TGFα and EGF Receptor in Mammary Gland Development and Neoplasia

Transforming growth factor-alpha (TGF)⁴ and/or the EGF receptor (EGFR) are frequently overexpressed by human and rodent breast tumors, as well as tumor-derived cell lines. Additionally, various observations suggest a role for TGF and the EGFR signaling system in normal mouse mammary gland development. Recently, several laboratories have established TGF transgenic mice with which to study the role of this growth factor in normal and neoplastic mammary biology. Examination of these mice revealed that overexpression of TGF has profound consequences for this tissue. Most strikingly, transgenic mice expressing TGF under the control of tissue-specific and nonspecific promoters stochastically developed focal mammary tumors with an incidence and latency that was markedly affected by pregnancy. Most TGF-induced tumors were well-differentiated adenomas/adenocarcinomas, although some were undifferentiated and locally invasive. Distant metastases were only occasionally observed. Administration of the genotoxic carcinogen, 7,12-dimethylbenzanthracene (DMBA), dramatically accelerated mammary tumorigenesis induced by the TGF transgene, raising the possibility that TGF acts as a promoter in this tissue. Mice harboring dual transgenes encoding TGF and either wild-type ERBB2 or c-myc displayed markedly accelerated tumorigenesis compared to mice carrying any of the single transgenes alone, indicative of potent cooperativity. Moreover, tumorigenesis in the bitransgenic mice was less dependent on pregnancy, and tumors were generally more malignant in appearance. Finally, TGF also affected mammary gland dynamics. TGF transgenic mice consistently displayed precocious alveolar development, were variably impaired with respect to lactation, and showed markedly reduced postlactional involution. As a result, the glands of multiparous females accumulated hyperplastic lesions that generally resembled milk-producing alveoli. Limited data support the hypothesis that these lesions were precursors to TGF-induced tumors. In summary, these various findings underscore the potential importance of TGF for cellular differentiation and transformation in the mammary gland. They also establish TGF transgenic mice as a powerful model with which to study the role of EGFR signaling molecules in this dynamic tissue.