Measurement of antinuclear antibodies: assessment of different test systems.

The performance of rat liver and HEp-2 in the detection of antinuclear antibodies (ANA) was studied by two independent sites and compared against an ANA enzyme immunoassay (EIA) screen and EIA systems for the measurement of antibodies to double-stranded DNA (dsDNA) and ENA. Sixty-two sera from patients with connective tissue disease (CTD) and 398 from controls suffering from other disorders were included. The level of agreement was, for HEp-2 and rat liver (within one site), 82.0% (ANA positive/ANA negative) and 51.0% (ANA pattern); and for HEp2- and HEp-2 (between sites), 71.8 and 86.5%. On sera with the ANA homogeneous pattern, the measurement of anti-ENA EIA added little to the detection rate with anti-dsDNA EIA alone. On ANA speckled sera, the EIA reactivity depended on the reaction of the mitotic cells: while sera with positive mitoses reacted similarly to ANA homogeneous sera, in those with negative mitoses the measurement of anti-ENA added about 10% to the detection rate achieved with anti-dsDNA alone. The measurement of anti-Scl-70 and anti-Jo-1 did not markedly improve the positive rate with classical ENA (anti-SSA, -SSB, -Sm, and -RNP) alone, raising doubts about the cost efficiency of including these measurements in unselected sera. The ANA EIA identified patients with CTD at a rate similar to that for rat liver and HEp-2. However, up to 98% of the sera found to be negative by ANA EIA but positive by use of rat liver and HEp-2 were from controls. Thus, the ANA EIA may possible be used as an alternative screen, particularly in laboratories with a high frequency of sera from patients not suffering from CTD.

Urinary alanine aminopeptidase and beta 2-microglobulin as measurements of aminoglycoside-associated renal impairment.

The value of urinary alanine aminopeptidase (AAP) and urinary beta 2-microglobulin as predictors of aminoglycoside-associated nephrotoxicity was studied in 46 patients treated with gentamicin or tobramycin. In three patients serum creatinine increased by more than 50 mumol/l. Urinary AAP increased in virtually all patients. The degree of these increases could not be correlated to subsequent increases in serum creatinine. Increases in urinary beta 2-microglobulin were also seen in many patients who did not show subsequent increases in serum creatinine. Moreover, urinary beta 2-microglobulin was elevated before the onset of aminoglycoside treatment in many patients with septicaemia and malignant diseases, thus making an evaluation of antibiotic-induced changes impossible. These results indicate that neither urinary AAP nor urinary beta 2-microglobulin can be used to predict aminoglycoside-associated nephrotoxicity of clinical importance in individual patients.

Renal function of neonates during gentamicin treatment.

Thirteen newborn infants, 8 term and 5 preterm (gestational age 31 to 36 weeks), were treated for between 3 and 7 days with gentamicin and ampicillin or cloxacillin because of suspected bacterial infection. The dosage of gentamicin was carefully monitored by serum concentration assays. Urinary alanine aminopeptidase, urinary beta 2-microglobulin, serum urea, and serum beta 2-microglobulin were measured during and after the end of treatment to detect signs of renal toxicity. Levels of urinary aminopeptidase increased in 12 of them, indicating damage to the cells of the proximal tubuli. Changes in urinary beta 2-microglobulin followed the normal physiological course seen in neonates after birth. Serum levels of urea and beta 2-microglobulin did not indicate any drug-associated depression of glomerular filtration rate.

Smoking and circulating IgE in bronchial carcinoma.

The serum concentrations of total IgE were significantly raised in smokers compared to those who had never smoked (p less than 0.005) among male patients with bronchial carcinoma, while no differences were found between smoking and non-smoking female bronchial carcinoma patients. The total IgE levels in male and female patients with non-malignant pulmonary diseases were not correlated to smoking habits. No significant differences in the IgE levels were observed between smoking males subgrouped according to the WHO histological types of bronchial carcinoma. Males with carcinoma who had stopped smoking more than 10 years ago had significantly reduced IgE levels compared to male cancer patients continuously smoking (p less than 0.01). These data, indicating that smoking is associated with elevated IgE levels in males with bronchial carcinoma, might suggest that smoking in certain, preferably male, individuals induces an impaired cellular immunity which is reflected by an enhanced IgE synthesis and a depressed resistance to carcinogens of tobacco smoke.

Comparison of serum beta 2-microglobulin and carcinoembryonic antigen (CEA) in the follow-up of breast cancer patients.

Using commercially available radioimmune test kits, serial determinations of serum beta 2-microglobulin and CEA were performed in 337 patients, who had been treated for breast cancer by modified radical mastectomy and radiotherapy. The pre-therapeutic data indicated a higher incidence of pathological beta 2-microglobulin and CEA levels in patients with distant metastases than in patients with localized disease. However, this finding did not allow the conclusion of a direct complementarity of beta 2-microglobulin and CEA as tumour markers, since the group of patients with distant metastasis contained a high percentage of elderly patients who generally can be expected to have elevated beta 2-microglobulin serum concentrations. Therefore, the correlation of the clinical course of malignant disease and the incidence of relapses with the changes of serum beta 2-microglobulin and CEA concentrations was examined during the post-treatment surveillance: 7/9 cases (78%) with local recurrence and 46/73 cases (63%) with distant spread of disease were not indicated in the beta 2-microglobulin follow-up by pathologic serum concentrations, whereas in the CEA follow-up only 1/9 and 2/73 false negative indications were registered. The poor correlation suggests that serum beta 2-microglobulin is not directly tumour associated in breast cancer and does not fulfill the criteria of a tumour marker.

Elevated serum immunoglobulin E in bronchial carcinoma: its relation to the histology and prognosis of the cancer.

Total IgE and specific IgE antibodies against six common allergens were measured in the sera of 217 unselected patients with bronchial carcinoma. Their median total IgE level was significantly increased as compared to the median levels found in two control populations consisting of 246 individuals representing the adult general population and 143 patients with benign pulmonary disorders. The frequency of serological atopy, i.e., the presence of specific IgE antibodies, was also significantly increased in the cancer population as compared to the controls. In contrast, the incidence of possible clinical atopy was about five times higher in the general population than in the cancer group. Patients with bronchial carcinoma typed as adenocarcinoma had the best prognosis and also had nonelevated IgE levels in contrast to patients with bronchial carcinoma typed as squamous cell carcinoma, small or large cell carcinoma. The favourable prognosis with nonelevated IgE levels also was demonstrated in patients with squamous cell carcinoma. It is suggested that the elevated IgE levels in bronchial carcinoma reflect impaired cellular immunity.

The self-sustaining capacity of lymphoid cells in the irradiated thymus.

The self-sustaining capacity of thymus lymphoid cells was studied in mice exposed to sublethal doses of X-rays. During exposure various parts of the organism were protected by lead shielding. When the thymus was protected and the rest of the organism exposed to radiation an initial cellular depletion of the thymus not exceeding 70 per cent was followed by a complete cellular regeneration of the organ within 3 weeks. This regeneration was shown to proceed without any cellular support from the bone marrow. When the thymus was included in the radiation field and/or damaged by cortisone treatment resulting in a depopulation exceeding 70 per cent, no or only partial regeneration occurred. The results indicate that the cell population in the thymus of mice can be self-sustaining for at least 3 weeks.

Is serum beta 2-microglobulin a tumor marker in gastrointestinal cancer?

Serial determinations of serum beta 2-microglobulin (beta 2m) and carcinoembryogenic antigen (CEA) were performed in 314 patients with histologically confirmed gastrointestinal cancer. The data were correlated with a set of clinical parameters. Pre-operative serum beta 2m levels did not discriminate different classes of tumor extension nor different stages of resectability of tumors in contrast to CEA. During post-operative surveillance the correlation of the time courses of serum beta 2m and CEA with the clinical course of malignant disease was studied in a selected group of 165 patients with resected primary carcinoma of the gastrointestinal tract. During the follow-up 74/165 patients showed disease progression or recurrence. In the beta 2m follow-up 66% false negative indications (49/74) of malignant disease were observed, whereas in the CEA follow-up it was 5% (4/74). The ratio of correct positive/false positive indications was 25/10 in the beta 2m follow-up and 70/10 in the CEA follow-up. The data indicate that the formation of serum beta 2m is not directly tumor associated in gastrointestinal cancer.

Effect of cortisone and X-irradiation on cellular depletion and regeneration in the thymus of mice: experimental discrimination between thymus lymphocyte precursors in the bone marrow and in the thymus.

The effect of cortisone treatment on the ability of bone marrow cells to repopulate X-irradiated thymus was investigated. In one experimental series, groups of mice were treated first with cortisone and then irradiated with or without bone marrow protection. Mice treated with either cortisone or radiation alone served as controls. During an initial, bone marrow independent phase of thymus regeneration, cortisone had a stronger inhibitory effect on the cellular regeneration of the organ than irradiation. On the other hand, during a subsequent bone marrow dependent phase, thymus regeneration was impaired by radiation exposure but not by cortisone treatment. In another experimental series, irradiated mice were transplanted with bone marrow cells in different numbers from syngeneic donors which had either been treated with cortisone or were left untreated. Twenty days later the cell number was consistently larger in the thymus of animals which has been transplanted with cortisone treated bone marrow than in the animals transplanted with untreated bone marrow. It is concluded that the thymus lymphocyte precursors in the bone marrow and the early precursors of thymocytes in the thymus differ with regard to their sensitivity to cortisone and radiation and, therefore, may represent two distinct cell types.

Dose to the shielded thymic region of irradiated mouse cadavers.

Frozen mouse cadavers were exposed to 220 kV roentgen irradiation with the thymic area covered with a lead plate sized 18 mm X 24 mm X 4 mm oe 18 mm X 12 mm X 4 mm. Lif-Telfon rods were used for dose measurements. With larger plate the mean dose from scattered radiation, in percent of that in the uprotected abdomen, was 12.77 (thymic lobes) and 18.34 (vertebra). With the smaller shield the figures were markedly increased, 16.70 and 23.77, respectively. Since the percentage dose is relatively high, the amount of scattered radiation may have biologic significance.

Cellular repopulation and recovery of the phytogemagglutinine (PHA) responding cell pool in the thymus of sublethally irradiated mice.

Cellular repopulation and recovery of PHA reactivity was investigated in the thymus of mice exposed to 400 R either on the whole body or with one leg protected. In the whole body irradiated cases both the cellular repopulation and the restoration of PHA reactivity in the thymus showed a cyclic pattern: an initial depression and a subsequent first recovery being followed by a secondary cellular depletion and decrease of reactivity before another recovery phase started. The phase of the secondary decrease lasted fo a similar period for both the cell number and reactivity, but was shifted in time and the reduction in reactivity began some 12 days later than that in the cell number. In the leg-shielded animals, the cellular repopulation progressed without cyclic changes, but the recovery of PHA reactivity maintained a cyclic pattern. However, due to a smaller secondary decrease, it was less pronounced than in the whole body irradiated cases. During the aging of mice from 50 to 90 day old, the cell number in the involuting thymus decreased to half of the original. The decrease in the cell population was associated with an about four-fold increase of PHA reactivity. The results were discussed in regard to the origin of the immunoreactive thymic cells. Arguments were presented in favour of the hypothesis that both the reactive and nonreactive cells originate from precursors in common, but require different lengths of time for maturation.

The role of bone marrow in different phases of the cellular repopulation of irradiated mouse thymus.

In one series of experiments, the cellular repopulation of the thymus was investigated in mice exposed first to 200 R on the whole body and, after various intervals, to 700 R with one leg protected or unprotected during the exposure. When no protection was made, the mice were transplanted with syngeneic bone marrow cells in a defined number immediately after irradiation. Repopulation was fastest when the interval between exposures was 5 days, and most delayed when it was 14 days; with a 30 day interval the speed of repopulation was intermediate, and resembled that of a control group exposed to only the second dose. In another experimental series, thymus repopulation was studied after exposure of mice first to 200 R with one leg protected or unprotected and, after an interval of 5, 14 or 30 days, to a second dose of 700 R on the whole body. In all cases, syngeneic bone marrow cells were grafted intravenously after the second irradiation. The thymus repopulation was enhanced by protection of the leg when 14 day interval separated the exposures. In the other cases, no enhancement was noted. The findings were interpreted to indicate that the cellular composition of the thymus and, in particular, the frequency of the proliferating stem cells at the time of the exposure determines thymic repopulation for about two weeks after irradiation. After this period, repopulation is due to new precursors from the bone marrow which seeded the thymus.

[Influence of a sublethal irradiation on the immune response of the mouse to sheep erythrocytes]. / Influence d'une irradiation sublétale sur la réponse immunitaire de la souris aux hématies de mouton

In the CBA mice, the immunological response of the spleen cells (RFC and PFC direct and indirect) against the sheep erythrocytes is highly depressed by a 400 R dose of X rays. The recovery is not complete at the 30th day after irradiation. The response of the bone marrow cells either irradiated or unirradiated to the antigenic stimulation is very low.

[Immuno-cyto-adhesion and sublethal irradiation]. / Immuno-cyto-adhérence et irradiation sublétale

1. In sublethally irradiated CBA mice, the relative and absolute numbers of spontaneous rosetts forming cells against sheep erythrocytes are markedly decreased in bone marrow. 2. The decrease of the absolute number of spontaneous RFC is also important in the spleen in spite of an increase of the RFC relative number above the normal values between the 8th and 12th day after irradiation. 3. The graft of normal bone marrow cells immediately after irradiation of the shielding of a medullary area during irradiation promotes the recovery of the immunocytoadherence capacity of the bone marrow cells but not of the spleen cells.