Early Research Articles

Effects of immunotherapy on the early, late, and rechallenge nasal reaction to provocation with allergen: Changes in inflammatory mediators and cells

We investigated the effects of immunotherapy (IT) on the early (ER), late (LPR), and rechallenge reactions (RCRs) to nasal challenge with antigen as well as on the cutaneous ER and LPR to intradermal skin challenge. Our expectation was that IT would have a preferential effect on the LPR, and our aim was to understand the mechanism. Twenty-one ragweed hay fever-sensitive subjects were treated with a moderate dose of antigen extract (maintenance dose of 1.94 μg of antigen E ( Amb a I)) during a period of 8 months (total dose equivalent to 24 (μg of antigen E), and 20 matched subjects received placebo injections in a double-blind manner. Both groups underwent identical nasal challenges and intradermal skin tests with ragweed-antigen extract both before 1985 and during 1986 IT. Symptom and medication diaries, recorded during seasonal exposure, and changes in specific serum IgE and IgG antibodies confirmed the efficacy of the administered IT dose. Between-group analysis revealed that IT significantly reduced the levels of histamine, TAME-esterase activity, and kinins, as well as symptoms of rhinorrhea and congestion generated during the ER to nasal challenge. Within-group paired analysis demonstrated ER, LPR, and RCR mediators and symptoms also to be reduced by IT. Surprisingly, the placebo-treated group demonstrated an increase in the ER. There was no decrease of the LPR without an antecedent decrease of the ER. IT did not clearly change the late cellular inflammatory response. In the case of skin challenge, IT significantly reduced the cutaneous ER. The reduction of the cutaneous LPR was more pronounced. We speculate that moderate-dose IT ameliorates seasonal symptoms of allergic rhinitis by reducing the ER, LPR, and RCR to antigen challenge but does not preferentially reduce the nasal LPR.

Analysis of the forelimb crossed extension reflex in thalamic cats during stepping

Forelimb crossed extension reflexes were examined in 22 thalamic cats. These reflexes were elicited either by backward passive movement or by repetitive electrical stimulation of cutaneous and joint afferent nerves in the contralateral forelimb. Single stimulation of the superficial radial nerve evoked two types of reflex responses—early (ER) and late (LR)—from the triceps brachii muscle on the contralateral side. The latencies were about 7 and 16–25 ms, corresponding to the propriospinal (PSR) and spino-bulbo-spinal (SBS) reflexes of the ipsilateral flexor, respectively. Repetitive stimulation of the superficial radial nerve evoked the LR but not the ER. The crossed extension reflex and LR were abolished by lesions of the dorsolateral funiculus of the cervical cord on the side opposite to the recording. The tonic EMG activity, crossed extension reflex and LR in the extensor on the side of lesions were abolished by lesions of the ventrolateral funiculus of the cervical cord. During forelimb stepping, the amplitudes of both ER and LR fluctuated depending on the phase of the step cycle. The ER appeared during a narrow period in the early phase of the stance, whereas the LR was observed during a wide period from the middle of the swing to the middle of the stance. Both responses were absent from the middle of the stance to the middle of the swing. These observations suggest that forelimb crossed extension reflexes involve both spinal and supraspinal (SBS) loop mechanisms, and that these are utilized during stepping, with the latter mechanism in particular playing an important part in the extension phase of the forelimb forward movement.

Plasma growth hormone and insulin concentrations in, and free fatty acid release from adipose tissue cultured in vitro from Holstein cows of differing cow index during early and late lactation

Early (EL) and late (LL) lactation Holstein cows were segregated into three Cow Index (CI) groups (high, HG; medium, MG; low, LG; n=47). Feed intake by lactation group, individual milk yield data and blood samples, obtained by puncture of the coccygeal vein or artery at 12-hr intervals, were collected for 7 d. Cows were fed alfalfa hay top dressed with grain mixture. On day 7, 5 g of subcutaneous adipose tissue were removed from the tail-head region. Tissue was minced into 10–15 mg pieces in Krebs-Ringer bicarbonate buffer with 5 ng/ml insulin added (KRB). Triplicate 100-mg aliquots were incubated in KRB + 3% essentially fatty-acid-free bovine serum albumin with either 50 ng/ml growth hormone (G), 5 μg/ml epinephrine (EPI), both (G+E) or neither (CON) at 37 C for 2 hr. Early lactation cows averaged greater (P<.05) daily milk production (33.4 vs 22.1 kg), greater (P<.05) plasma growth hormone (GH) concentrations (3.9 vs 3.0 ng/ml) but lesser (P<.01) insulin (INS) concentrations (.49 vs .73 ng/ml) than LL cows. Adipose tissue FFA release in vitro was greater (P<.01) when media contained EPI (EPI: 8.10; G+E: 8.05 μE/l/g tissue) than when EPI was not present (CON: 1.33; G: 1.39 μE/l/g tissue), but was not affected by stage of lactation. Including hormonal data in the model as covariates indicated that increased plasma INS concentrations before biopsy reduced subsequent FFA release in vitro when tissue was incubated with added EPI, but not when incubation media lacked EPI. Increased GH concentrations had the opposite effect. Further, FFA release was greatest from HG cow adipose when incubated in media lacking EPI, but greatest from LG cow adipose when incubated in media containing EPI.

Relationship between the early, late, and rechallenge reaction to nasal challenge with antigen: Observations on the role of inflammatory mediators and cells

We challenge each of 55 consecutive ragweed (RW)-allergic patients with hay fever and with graded increasing doses of ragweed extract to investigate the frequency and relationship between the early (ER), late (LPR), and rechallenge reactions (RCRs) to nasal challenge. We evaluated the nasal response by measuring the levels of histamine, TAME-esterase activity, and kinins in the nasal lavage fluid and by grading symptoms. Fifty-one subjects (92.7%) had an ER consisting of a dose-dependent, concommitant increase in both mediators and symptoms. The total amount of TAME-esterase activity and kinins generated during ER correlated significantly with specific serum IgE (ssIgE), intradermal skin test (ST) sensitivity, and basophil histamine release (BHR) to antigen E (p less than 0.01 for each). Twenty-four (47%) subjects developed a late increase in mediators and 23 (45%) subjects in symptoms. None of the four subjects without an ER developed an LPR. The levels of the late-appearing mediators were not predicted by ST, ssIgE, or BHR. There was a significant but weak association between the intensity of ER and LPR, but there was no significant difference in the IgE antibodies, ST, BHR, and intensity or threshold of ER between dual and early only reactors. The number of eosinophils and neutrophils in the LPR lavages increased over the prechallenge baseline, and their numbers correlated (p less than 0.05) with ER kinins (r = 0.46, and 0.37, respectively), ER TAME-esterase activity (r = 0.28 and 0.24, respectively), and in the case of eosinophils, ER histamine (r = 0.29).(ABSTRACT TRUNCATED AT 250 WORDS)

Effects of induced parturition and early obstetrical assistance in beef cattle.

Pregnant crossbred beef females (33 second-calf cows and 73 primiparous heifers) bred to a single Hereford sire were assigned to a 2(3) factorial study involving age of dam, natural (NP) or induced (IP) parturition and late emergency (LA) or forced early (EA) obstetrical assistance. Parturition was induced with 10 mg flumethazone given i.m. between 1400 and 1600 on d 272 of gestation; EA was given when the cervix and birth canal were fully dilated. Average IP occurred 39.6 h postinjection, and 95.3% of the treated dams responded within 60 h postinjection; gestation was shortened 2.9 d (P approximately equal to .07). Dystocia score (from 1 = no assist to 4 = major traction required and 5 = abnormal presentation) was 1.12 vs 2.40 for LA and EA, respectively (P less than .01), and 11% of LA vs 84% of EA were assisted. Calf vigor score (1 = normal to 3 = severely depressed or dying) at birth was 1.3 for NP and 1.1 for IP (P approximately equal to .06) and 1.3 for EA and 1.1 for LA (P less than .05). This effect of EA was due to reduced vigor of calves experiencing abnormal presentation. Birth weights (BW) and weaning weights (WW) of calves from cows exceeded those from heifers (32.6 vs 30.8 kg, P less than .05; 210.9 vs 156.3 kg, P less than .01, respectively). Differences due to IP and EA in BW, WW, postpartum interval and conception rate were not significant, but weight gain of calves from EA dams tended (P approximately equal to .09) to be greater than weight gain of calves from LA dams.(ABSTRACT TRUNCATED AT 250 WORDS)

Early interactions between human cytomegalovirus and cells.

Adsorption of cytomegalovirus (CMV) to human fibroblasts was not inhibited by preincubation with other herpes viruses (HSV-1, HSV-2, VZV). Transport of virus to the nucleus was studied using virus labelled with 3H-thymidine. Radioactivity was found in the nucleus 20 minutes after virus had been added to the cells. In order to assess the expression of the virus genome, synthesis of early (EA) and late (LA) antigens was studied. Assayed on permissive human lung fibroblasts, a plaque purified virus contained more EA inducing than both EA and LA inducing viral units. Since this was a consistent finding with virions of all densities, it seems to be an effect of viral dose rather than of virion density. Alternatively, LA-defectiveness is a virion property which does not vary with virion density.

Infection of the human T-cell-derived leukemia line Molt-4 by Epstein-Barr virus (EBV): Induction of EBV-determined antigens and virus reproduction

We have reported previously that human T-cell-derived Molt-4 cells become susceptible to Epstein-Barr virus (EBV) infection after implantation of functional EBV receptors into the plasma membranes of Molt-4 cells (Volsky et al., 1980). In the present work, we expand this finding by analyzing the following: (i) Virus adsorption vs viral penetration—using [ 3H]thymidine-labeled EBV, we demonstrate that the virus could adsorb to both the untreated and the receptor-implanted Molt-4 cells. However, only the altered cells became susceptible to EBV penetration followed by the viral antigen synthesis; (ii) EBV substrain specificity of infection—EBV P3HR-1 virus induced the nuclear (EBNA), early (EA), and virus capsid (VCA) antigens, whereas EBV B-95-8 virus induced only EBNA; (iii) virus reproduction— in situ hybridization was used to demonstrate the EBV-DNA synthesis in the P3HR-1 virus-infected cells. In addition, immature herpes-like particles were observed in electron micrographs of infected cells. It is concluded that EBV infection of human T-cell-derived Molt-4 cells may lead to a full viral-lytic cycle in a portion of infected cells. The results suggest, however, that the primary EBV productive infection may not necessarily involve any immunofluorescence-detectable EBNA synthesis.

Activation of Epstein-Barr virus expression in human lymphoblastoid P3HR-1 and Raji cells with propionic acid and with culture fluids of propionic acid-producing anaerobes

Epstein-Barr virus (EBV)-associated early (EA) and virus capsid antigens (VCA) were efficiently induced in the viral genome-carrying human lymphoblastoid cells, P3HR-1 and Raji, by the culture fluids of Propionibacterium acnes, P. avidum, P. lymphophilum and Arachnia propionica, the anaerobes which are commonly seen among the normal flora of man. The active principle for EBV-induction in the 2 cell lines was the propionic acid produced by the microbes and such activity was shown to correlate with the fatty acid content of the culture media.

Combined effect of the extracts from Croton tiglium, Euphorbia lathyris or Euphorbia tirucalli and n-butyrate on Epstein-Barr virus expression in human lymphoblastoid P3HR-1 and Raji cells

The combined usage of n-butyrate and 12- O-tetradecanoylphorbol-13-acetate (TPA) or the oily extracts from Croton tiglium, Euphorbia lathyris or Euphorbia tirucalli exerted a marked effect on induction of Epstein-Barr virus (EBV)-associated early (EA) and viral capsid (VCA) antigens in EBV genome-carrying human lymphoblastoid cell lines. In producer P3HR-1 cells, the enhancing effect of the 2 components was additive both for EA and VCA, while in non-producer Raji cells, a synergistic increase of EA was observed. The possible implication of these findings relating to the cause of EBV-associated diseases is discussed.

Productive Epstein‐Barr viral infection of the human lymphoblastoid cell line 6410 with release of early antigen inducing and transforming virus

The human lymphoblastoid cell line 6410 was superinfected with P3HR-1 derived Epstein-Barr virus. Subsequent to the first cycle of infection, characterized by early (EA) and virus capsid (VCA) antigen synthesis, the culture, designated 6410-EBV, continued to synthesize VCA. Further immunofluorescence and electron microscopic examination over 18-24 months showed the 6410-EBV culture to be productively infected with EBV. Virus was recovered, in quantity, from the culture fluids and assayed for ability to induce EA synthesis in Raji cells and to transform human umbilical cord lymphocytes. The virus was found to possess both properties, in contrast to P3HR-1 virus which only induces EA. HLA and karyologic analyses of P3HR-1, 6410 and 6410-EBV cultures showed relatedness of 6410-EBV to 6410 cells and dissimilarity to P3HR-1. The biologic activity data coupled with the cytologic analyses confirm a productive infection of the target cells. The reason for differences in biologic activity between the infecting (P3HR-1) and progeny virus is unresolved. It may be speculated that the endogenous EBV genome of 6410 cells was activated and gave rise to a different strain of EBV or to a mixed progeny-parent population as a result of dual infection. Alternatively, the parent strain of virus may have been modfied as a result of interaction with the new host cell.

Complexity of EBV homologous DNA in continuous lymphoblastoid cell lines

Abstract The complexity of Epstein-Barr Virus (EBV) homologous DNA in 11 EBV-infected lymphoblastoid cell lines which have been passaged for several years in culture was determined by hybridization of lymphoblastoid cell DNA to DNA extracted from EBV purified from HR-1 cells and labeled in vitro. Five of the cell lines analyzed contained no early (EA) or viral capsid (VCA) antigens which have been associated with the replication of EBV. Two other cell lines contained EA but not VCA. Of the seven VCA-negative cell lines, those which contained some early antigen or in which early antigen could be induced with IUDR had 56, 48, and 25 copies per diploid cell genome of more than 90% of the sequences of EBV DNA. Five cell lines which did not contain EA even after induction had 23, 8, 8, 6, and 2 copies per cell of the EBV genome. The data indicate that there is a correlation between the number of copies of EBV DNA in nonpermissive cells and the ability of these cells to express EA. Three of the five EA and VCA negative nonpermissive cell lines contained DNA homologous to more than 90% of the sequences of EBV DNA. The kinetics of hybridization of the DNA of two other nonpermissive cell lines, Namalwa and SKL, which contain two and eight copies, respectively, of some EBV DNA sequences, suggest but do not prove that these cell lines may contain incomplete viral genomes. The retention of the full complexity of viral DNA in most nonpermissive lymphoblastoid cell lines may be related to the relatively large number of copies of the viral genome in these cells.