Aimée Reuter

Tumor necrosis factor and interleukin-1 serum levels during severe sepsis in humans.

In a study of serum levels of tumor necrosis factor (TNF alpha) and interleukin-1 beta (IL-1 beta) in patients developing sepsis in the ICU, high TNF alpha levels were found in patients with septic shock. Normal values are 75 +/- 15 pg/ml; in these patients, TNF alpha serum level ranged from 100 to 5000 pg/ml with a mean of 701 +/- 339 pg/ml and a median of 250 pg/ml. There was a correlation between TNF alpha level and sepsis severity score as well as with mortality. In contrast, IL-1 beta serum levels were only slightly increased and were not correlated with severity or mortality.

IN VIVO CELL ACTIVATION FOLLOWING OKT3 ADMINISTRATION: SYSTEMIC CYTOKINE RELEASE AND MODULATION BY CORTICOSTEROIDS

A massive and self-limited release of tumor necrosis factor and interferon gamma was detected in the systemic circulation in 35 consecutive renal allograft recipients by specific radioimmunoassays very soon following the first injection of the monoclonal antibody OKT3 (anti-CD3). Peak serum TNF and IFNγ levels were reached, respectively, at 1 and 4 hr following the first OKT3 injection. Abnormally high serum interleukin 2 levels were also observed 4 hr following the first OKT3 injection in a minority of patients (5 cases). OKT3 had no effect on interleukin 1 beta, interferon alpha, and granulocyte/macrophage colony stimulating factor serum levels, which in all patients remained within the normal range throughout the study. This selective OKT3-induced cytokine release, which only followed the first injection, was transient (i.e., lasting a few hours). It tightly paralleled the spontaneously reversible clinical syndrome characterized by high fever, headaches, and gastrointestinal symptoms that is invariably associated with the first OKT3 administration. Importantly, when administered in adequate dosages and with adequate timing, corticosteroids influenced both the cytokine release and the systemic reaction. Thus, the highest TNF, IFNγ, and IL-2 serum levels were detected in patients who did not received corticosteroids. Patients who received high-dose corticosteroids (1 g solumedrol bolus) concomitantly with the first OKT3 injection still had high TNF and IFNγ levels. Conversely, when the same corticosteroid dose was injected 15–60 min prior to the first OKT3 injection, in all cases the increase of serum TNF and IFNγ was significantly lower as compared with the above-described groups; IL-2 levels did not rise. These data offer a direct explanation for one major side effect of OKT3 and thus provide the basis for devising means to prevent its occurrence.

POLYMORPHISM OF PROTEIN AND POLYPEPTIDE HORMONES

Work of the past few years indicates that many of the protein and polypeptide hormones exist in several forms both in their gland of origin and in the plasma. This heterogeneity was established by radioimmunological studies of these hormones in different biological fluids or in the gland of origin and by chemical characterization of these hormones after purification. For example, the heterogeneity of parathyroid hormone (PTH) was first described by Berson & Yalow (1968) who noted a dissociation between the immunological activity of plasma PTH and the hormone in gland extracts with one antiserum, whereas with another antiserum there was no difference between the immunological behaviour of these two PTH-containing media. This indicated that the first antiserum contained antibodies against antigenic groups present in the molecule of PTH extracted from the gland but absent, or modified in their binding affinity, in the molecule of plasma PTH. The other antiserum contained only antibodies directed against antigenic groups present on the PTH molecules both in plasma and in the gland extract. Striking examples of heterogeneity of plasma hormone and hormone in glandular extracts have been uncovered in the case of insulin, ACTH, PTH, thyrocalcitonin, gonadotrophins, glucagon, TSH, gastrin and perhaps growth hormone. This heterogeneity of protein and polypeptide hormones may be due to one or more of the causes listed in Table 1. The theoretical possibility of carrier protein is represented on the table but it has not been established that any circulating peptide hormone is bound to serum proteins. We should like to discuss some of these causes of heterogeneity and analyse the evidence that would allow us to conclude that this hormonal polymorphism is either biologically significant or the result of chemical artefacts.

Homologous radioimmunoassay for human prolactin

Radioimmunoassays for human prolactin in various biological fluids have been available since 1971.(1–7) Most of these assays have used heterologous systems with prolactin preparations from various sources, i.e. sheep, pig and primate with their corresponding antiserum.(1–5)Friesenet al.(6) find Sinhaet al.(7) are the only authors who have described homologous assays using human prolactin as the reference standard and tracer as well and an anti-human prolactin antiserum. In the present study we have established a homologous radioimmunoassay using human pituitary prolactin. The separation method is based on the double antibody solid phase system.(8) Cross reactivity with human growth hormone (HGH), placental lactogen (HPL), the pituitary protein hormones and prolactins of various species were studied as were values found in normal subjects in basal conditions and after a TRH injection. The results of measuring basal levels and performing dynamic tests under various physiological and pathological circumstances will be reported subsequently.

PROLACTIN LEVELS DURING THE MENSTRUAL CYCLE

The levels of prolactin, FSH, LH, oestradiol and progesterone were measured daily during fourteen ovulatory cycles. The behaviour of FSH, LH, oestradiol and progesterone was classical. Non‐systematic changes occurred in prolactin levels during the course of the menstrual cycle with the highest level being either during the ovulatory period or during the luteal phase. However, the mean level of prolactin was significantly higher during the ovulatory and luteal phases than during the follicular phase. A direct relationship between oestradiol and prolactin levels was noted, although there was no correlation between prolactin on the one hand and FSH, LH and progesterone on the other.

SERUM CONCENTRATIONS OF HUMAN CHORIONIC GONADOTROPHIN AND ITS ALPHA AND BETA SUBUNITS. 1. DURING NORMAL SINGLETON AND TWIN PREGNANCIES

Homologous radioimmunoassays have been developed for native hCG and its α and β subunits. Their specificities were assessed by analysis of the inhibition curves and by gel filtration of purified preparations of native hCG, hCGα and hCGβ, and of sera from pregnant women, on Sephadex G 100. Gel filtration data indicated a true cross‐reaction of native hCG in the hCGβ and hCGα assays of 0.5 and 0.35%, respectively. After validation of the RIA procedures for unfiltered serum assays, we have demonstrated that in 311 maternal sera obtained in 207 normal singleton pregnancies, free circulating hCGα increases continuously, whereas free hCGβ and native hCG peak between the 8th‐10th week. The ratio (hCGβ/hCG) x 100 is greatest during the first trimester (2–3%) and decreases thereafter to around 1%. The ratio (hCGα/hCG) x 100 is < 1% at the beginning of pregnancy, 2% at 14 weeks and 15–20% from the 23rd week onwards. The levels of hCG and its subunits are higher in fifty‐two samples obtained throughout normal twin pregnancies than in corresponding single pregnancies, but the ratios subunit: hCG are the same, suggesting qualitatively similar placental secretion in both types of pregnancy. Analysis of pregnancy sera by gel filtration and assay of hCG and subunits in eluant fractions confirmed that free α and β subunits are present in the circulation and that hCGα rises while hCG and hCGβ fall in late pregnancy. Heterogeneity of the circulating hCGβ subunit is also demonstrated.

Simultaneous assays of cancer associated antigens in benign and malignant breast diseases.

Five tumor markers can be simultaneously determined in the serum by radioimmunoassay: carcino‐embryonal antigen (CEA), α‐fetoprotein (αFP), human chorionic gonadotrophin (HCG), β‐subunit of HCG (βHCG) and K‐Casein. In a series of 935 healthy subjects, these antigens remain undetectable or are detected within very precise limits. At the start of the clinical evolution of breast cancer, the incidence of pathological concentrations (i.e. increased as compared with the highest level observed in normal subjects) of at least one antigen is 69% in stages T1, T2, T3, N0, N1, M0. This high incidence is mainly due to a concomitant determination of CEA, K‐casein, HCG and β‐HCG. The α‐FP test is never positive, while the k‐casein concentration is particularly high in the first clinical stages of breast cancer and with metastases. The concomitant determination of these tumor markers may be a biological element contributing to the diagnosis of neoplasia, although it is neither an absolute nor a specific criterion. Indeed, a pathological concentration of at least one antigen was observed in 5.5% of the subjects presenting with benign mastopathy. When metastases occur (25 patients), the incidence of pathological concentrations of at least one antigen increases: at 88%, the absolute values of these levels are increasing simultaneously. The determination of the antigen concentration therefore allows an evaluation of the extension of the disease. Surgical removal reduces the incidence of positivity of these antigens to 34%. Persistance of pathological levels seems to be related to a possibility of relapse or metastatic spreading. Finally, chemotherapy and radiotherapy applied on a tumor which is not excised, do not decrease the incidence of positivity of the tumoral markers, although their levels seem to fluctuate with the clinical evolution.

Clinical Use of OKT3: The Role of Cytokine Release and Xenosensitization

Publisher Summary This chapter presents the clinical use of OKT3. It discusses the role of cytokine release and xenosensitization. The clinical use of immunosuppressive anti-T cell murine monoclonal antibodies (MoAbs) has so far mostly been confined to transplantation programs. The chapter presents an indicative although non-exhaustive list of the different membrane molecules targeted by MoAbs used as immunosuppressors in transplantation. The MoAb that has been the most widely used in this setting is OKT3 (anti-CD3). OKT3 has been shown by different authors to be a very potent immunosuppressor able effectively to treat and also prevent acute allograft rejection. OKT3, thus, is a good example of a MoAb that in many transplantation centers is progressively replacing conventional serotherapy with polyclonal anti-lymphocyte sera. It has been a common experience of all centers using OKT3 that the first injections of MoAb invariably induce a flu-like syndrome, including in variable proportions depending on the patient, chills, headache, pyrexia, vomiting, diarrhea, tachycardia, respiratory distress, hypotension, and arthragia.

Serum concentration of human chorionic gonadotropin and its alpha and beta subunits. 2. Trophoblastic tumours.

Using specific homologous radioimmunoassays of native hCG and its α and β subunits, we determined the levels of these glycoproteins in unfiltered maternal blood serially obtained in five non‐invasive hydatidiform moles before and after evacuation. Some of these samples were assayed after gel filtration chromatography on Sephadex G 100. Twelve samples, obtained in cases of invasive trophoblastic tumour after ablative surgery and chemotherapy, were also assessed for their hCG, hCGα and hCGβ content. In unaborted moles, mean circulating levels of native hCG and free hCGβ were considerably increased (seven and thirteen times, respectively) as compared to normal pregnancies of the same age, whereas levels of free hCGα were either normal or slightly elevated. Chromatographic analyses of molar sera confirmed the presence of free circulating subunits, and separated hCGβ in its monomeric form from its higher molecular weight form, the latter being in greater quantity than in normal pregnancy sera. In contrast, the elution profile of serum native hCG was comparable in cases of normal and molar pregnancy. Successful curettage was accompanied by a return to normal levels of the native hCG and its α and β subunits in 40–90 days. Persistence of tumour tissue was indicated by a slight increase in levels of native hCG and the β subunit. Determination of α subunit level was less useful for the detection of any relapse.

Changes in Fetal and Maternal Blood Levels of Prolactin, Cortisol, and Cortisone during Eutocic and Dystocic Childbirth

The changes in blood levels of prolactin, total and free cortisol, and cortisone were studied and compared in 51 mother-infant pairs, 30 with eutocic delivery and 21 with dystocic delivery. Regardless of the type of delivery, the newborn at term showed significantly higher prolactin and cortisone serum levels than their mothers, and significantly lower levels of free and total cortisol. In fetal distress of short duration, free cortisol levels were significantly raised in both the mother and the child, while prolactin and cortisone levels were significantly higher only in the child. In contrast to these observations, serum prolactin and cortisone levels in the mother were not altered by the occurrence of fetal distress. In the newborn at delivery there was a negative correlation between serum prolactin and the Apgar score at 1 min applied to the part of the graph between 8 and 2 Apgar scores. This study illustrates the utility of fetal prolactin measurements in evaluating the stress to which the fetus is subjected.