STUDIES ON THE IMMUNOTHERAPY OF RUNT DISEASE IN RATS

Using rats of the Lewis and BN (Ag-B locus incompatible) isogenic strains, a comparative study has been made of the capacity to prevent or mitigate the development of runt disease with: (a) lymph node cell suspensions from normal adult BN rats, (b) node cells, or (c) serum from donors sensitized against Lewis tissue antigens, or (d) heterologous anti-lymphocyte serum (ALS) raised in rabbits against rat thymocytes. Following a standard intravenous or intraperitoneal inoculation of 20 x 106 Lewis node cells into neonatal BN hosts, there are cutaneous manifestations of runt disease within 11–15 days and death invariably takes place within 20 days. However, complete protection is afforded by administration of a similar number of normal BN node cells via a different vein, or admixed with the otherwise harmful Lewis node cells. However, timing of the administration was crucially important—precedence or delay by as little as 4 hr resulted in a great impairment of protection. When the inoculations of the two cell suspensions were separated by 24 hr, no protection was afforded. These and other observations suggested that a necessary condition for protection of the hosts by unsensitized isologous cells requires that they establish a prompt and intimate confrontation with the homologous target cells. At the same dosage level, suspensions of node cells from sensitized isologous donors were much more effective therapeutically, saving the lives of 92% of treated subjects when administered after a delay of 3 days, and of 19% when the delay was 4 or 5 days. Of the various immunotherapeutic agents studied, daily injections of 0.2 ml of isoantiserum gave the best results, and could totally reverse the course of the disease even when initiated at age 10–13 days and subjects already presented symptoms. ALS, although inferior to isoantiserum at the dosage levels tested, proved to be superior to sensitized isologous cells as a protective agent, since the initiation of daily injections after delays of 6 or 8 days were still effective. The observations that delayed treatments of infant rats with isoantisera or ALS resulted in complete recoveries sustain the thesis that the lesions responsible for the fatal outcome of runt diseases are not inflicted at a very early stage. The efficacy of both isoantisera and ALS as a means of inhibiting the progression of homologous disease also suggests that they may have therapeutic value in situations where this condition is encountered.

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Berberine and Obatoclax Inhibit SARS-Cov-2 Replication in Primary Human Nasal Epithelial Cells In Vitro

Viruses ◽

10.3390/v13020282 ◽

2021 ◽

Vol 13 (2) ◽

pp. 282

Author(s):

Finny S. Varghese ◽

Esther van Woudenbergh ◽

Gijs J. Overheul ◽

Marc J. Eleveld ◽

Lisa Kurver ◽

...

Keyword(s):

Epithelial Cells ◽

Early Stage ◽

Antiviral Drugs ◽

Target Cells ◽

Nasal Epithelial Cells ◽

Cytokines And Chemokines ◽

Human Nasal Epithelial Cells ◽

Apoptotic Protein ◽

Air Liquid Interface

Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) emerged as a new human pathogen in late 2019 and it has infected over 100 million people in less than a year. There is a clear need for effective antiviral drugs to complement current preventive measures, including vaccines. In this study, we demonstrate that berberine and obatoclax, two broad-spectrum antiviral compounds, are effective against multiple isolates of SARS-CoV-2. Berberine, a plant-derived alkaloid, inhibited SARS-CoV-2 at low micromolar concentrations and obatoclax, which was originally developed as an anti-apoptotic protein antagonist, was effective at sub-micromolar concentrations. Time-of-addition studies indicated that berberine acts on the late stage of the viral life cycle. In agreement, berberine mildly affected viral RNA synthesis, but it strongly reduced infectious viral titers, leading to an increase in the particle-to-pfu ratio. In contrast, obatoclax acted at the early stage of the infection, which is in line with its activity to neutralize the acidic environment in endosomes. We assessed infection of primary human nasal epithelial cells that were cultured on an air-liquid interface and found that SARS-CoV-2 infection induced and repressed expression of specific sets of cytokines and chemokines. Moreover, both obatoclax and berberine inhibited SARS-CoV-2 replication in these primary target cells. We propose berberine and obatoclax as potential antiviral drugs against SARS-CoV-2 that could be considered for further efficacy testing.

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INTESTINAL OBSTRUCTION

Journal of Experimental Medicine ◽

10.1084/jem.17.3.286 ◽

1913 ◽

Vol 17 (3) ◽

pp. 286-306 ◽

Cited By ~ 49

Author(s):

G. H. Whipple ◽

H. B. Stone ◽

B. M. Bernheim

Keyword(s):

Blood Pressure ◽

Intestinal Obstruction ◽

Intravenous Injection ◽

Intestinal Mucosa ◽

Closed Loop ◽

Fatal Outcome ◽

Toxic Substance ◽

Protective Agent ◽

Low Blood Pressure ◽

Closed Duodenal Loop

Closed duodenal loops may be made in dogs by ligatures placed just below the pancreatic duct and just beyond the duodenojejunal junction, together with a posterior gastro-enterostomy. These closed duodenal loop dogs die with symptoms like those of patients suffering from volvulus or high intestinal obstruction. This duodenal loop may simulate closely a volvulus in which there has been no vascular disturbance. Dogs with closed duodenal loops which have been washed out carefully survive a little longer on the average than animals with unwashed loops. The duration of life in the first instance is one to three days, with an average of about forty-eight hours. The dogs usually lose considerable fluid by vomiting and diarrhea. A weak pulse, low blood pressure and temperature are usually conspicuous in the last stages. Autopsy shows more or less splanchnic congestion which may be most marked in the mucosa of the upper small intestine. The peritoneum is usually clear and the closed loop may be distended with thin fluid, or collapsed, and contain only a small amount of pasty brown material. The mucosa of the loop may show ulceration and even perforation, but in the majority of cases it is intact and exhibits only a moderate congestion. Simple intestinal obstruction added to a closed duodenal loop does not modify the result in any manner, but it may hasten the fatal outcome. The liver plays no essential role as a protective agent against this poison, for a dog with an Eck fistula may live three days with a closed loop. A normal dog reacts to intraportal injection and to intravenous injection of the toxic substance in an identical manner. Drainage of this loop under certain conditions may not interfere with the general health over a period of weeks or months. Excision of the part of the duodenum included in this loop causes no disturbance. The material from the closed duodenal loops contains no bile, pancreatic juice, gastric juice, or split products from the food. It can be formed in no other way than by the activity of the intestinal mucosa and the growth of the intestinal bacteria. This material after dilution, autolysis, sterilization, and filtration produces a characteristic effect when introduced intravenously. When in toxic doses it causes a profound drop in blood pressure, general collapse, drop in temperature, salivation, vomiting, and profuse diarrhea, which is often blood-stained. Splanchnic congestion is the conspicuous feature at autopsy and shows especially in the villi of the duodenal and jejunal mucosæ. Adrenalin, during this period of low blood pressure and splanchnic congestion, will cause the usual reaction when given intravenously, but applied locally or given intravenously it causes no bleaching of the engorged intestinal mucosa. Secretin is not found in the duodenal loop fluid, and the loop material does not influence the pancreatic secretion. Intraportal injection of the toxic material gives a reaction similar to intravenous injection. Intraperitoneal and subcutaneous injections produce a relatively slow reaction which closely resembles the picture seen in the closed duodenal loop dog. In both cases there is a relatively slow absorption, but the splanchnic congestion and other findings, though less intense, are present in both groups. There seems, therefore, to be no escape from the conclusion that a poisonous substance is formed in this closed duodenal loop which is absorbed from it and causes intoxication and death. Injection of this toxic substance into a normal dog gives intoxication and a reaction more intense but similar to that developing in a closed-loop dog.

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RASSF1C oncogene elicits amoeboid invasion, cancer stemness and invasive EVs via a novel SRC/Rho axis

10.1101/2021.01.05.425393 ◽

2021 ◽

Author(s):

Maria Laura Tognoli ◽

Nikola Vlahov ◽

Sander Steenbeek ◽

Anna M. Grawenda ◽

Michael Eyres ◽

...

Keyword(s):

Breast Cancer ◽

Stem Cell ◽

Cancer Metastasis ◽

Early Stage ◽

Methylation Status ◽

Target Cells ◽

Stem Cell Markers ◽

Breast Cancer Patients ◽

Actomyosin Contractility ◽

Amoeboid Cells

AbstractCell plasticity is a crucial hallmark leading to cancer metastasis. Upregulation of Rho/ROCK pathway drives actomyosin contractility, protrusive forces and contributes to the occurrence of highly invasive amoeboid cells in tumors. Cancer stem cells are similarly associated with metastasis, but how these populations arise in tumors is not fully understood. Here we show that the novel oncogene RASSF1C drives mesenchymal to amoeboid transition and stem cell attributes in breast cancer cells. Mechanistically, RASSF1C activates Rho/ROCK via SRC mediated RhoGDI inhibition, resulting in generation of actomyosin contractility. Moreover, we demonstrate that amoeboid cells display the cancer stem cell markers CD133, ALDH1 and the pluripotent marker Nanog; are accompanied by higher invasive potential in vitro and in vivo; and employ extracellular vesicles to transfer the invasive phenotype to target cells and tissue. Importantly, the underlying RASSF1C driven biological processes concur to explain clinical data: namely, methylation of the RASSF1C promoter correlates with better survival in early stage breast cancer patients. Therefore, we propose the use of RASSF1 gene promoter methylation status as a biomarker for patient stratification.

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The Effects of Combined Insulin and Metformin Therapy in Obese Patients with Diabetes Mellits Type 2 in the Early Stage of the Disease

Bosnian Journal of Basic Medical Sciences ◽

10.17305/bjbms.2006.3173 ◽

2006 ◽

Vol 6 (2) ◽

pp. 54-58

Author(s):

Belma Aščić - Buturović

Keyword(s):

Insulin Therapy ◽

Pancreatic Beta Cells ◽

Early Stage ◽

Diabetes Type 2 ◽

Diabetes Type ◽

Target Cells ◽

Obese Patients ◽

Patients With Diabetes ◽

Insulin Activity

Combination of insulin and metformin has been shown to improve glycaemic control in clinical trials, particularly in obese patients with diabetes type 2. Insulin therapy can improve function of pancreatic beta cells and periphery insulin activity in target cells in order to enhance glycaemic homeostasis (1, 2, 3). In our study we included obese patients with diabetes type 2 in the early stage of the disease. The study is partially retrospective and partially prospective. The study encompassed 40 patients split in two groups. The first group of 20 patients received insulin therapy combined with metformin, while the patients of the second group were treated with oral antidiabetic drugs, sulfonylureas and metformin. Three months later, the group treated with insulin and metformin showed improvement in the monitored parameters, namely significant reduction in HbA1c (p = 0.003), MFBG (p = 0.0009), PPG (p = 0.028). Insulin therapy administered together with metformin, in obese patients with diabetes type 2, in the early stage of the disease, resulted in well regulated fasting blood glycaemia, as well as post challenge glycaemia and HbA1c.

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Target cells for granulocyte colony-stimulating factor, interleukin-3, and interleukin-5 in differentiation pathways of neutrophils and eosinophils

Blood ◽

10.1182/blood.v76.10.1956.1956 ◽

1990 ◽

Vol 76 (10) ◽

pp. 1956-1961 ◽

Cited By ~ 46

Author(s):

H Ema ◽

T Suda ◽

K Nagayoshi ◽

Y Miura ◽

CI Civin ◽

...

Keyword(s):

Bone Marrow Cells ◽

Early Stage ◽

Target Cells ◽

Granulocyte Colony Stimulating Factor ◽

Colony Stimulating Factor ◽

Interleukin 3 ◽

Cd34 Antigen ◽

Cd34 Cells ◽

Stimulating Factor

Abstract To study the relationship between hematopoietic factors and their responsive hematopoietic progenitors in the differentiation process, both purified factors and enriched progenitors are required. We isolated total CD34+ cells, CD34+,CD33+ cells, and CD34+,CD33- cells individually from normal human bone marrow cells by fluorescence- activated cell sorter (FACS), and examined the effects of granulocyte colony-stimulating factor (G-CSF), interleukin-3 (IL-3), and IL-5 on in vitro colony formation of these cells. CD34+,CD33+ cells formed granulocyte colonies in the presence of G-CSF. Both CD34+,CD33+ cells and CD34+,CD33- cells formed granulocyte/macrophage colonies in the presence of IL-3. Eosinophil (Eo) colonies were only formed by CD34+,CD33- cells in response to IL-3, but scarcely formed by CD34+ cells in the presence of IL-5. We performed the two-step cultures consisting of the primary liquid culture for 6 days and the secondary methylcellulose culture, and serially examined changes in phenotypes of ,he cells cultured in the primary culture. CD34-,CD33+ cells derived from CD34+,CD33+ cells by preincubation with G-CSF or IL-3 formed Eo colonies in the presence of IL-5 but not IL-3. CD34-,CD33+ cells derived from CD34+,CD33- cells by preincubation with IL-3 also formed Eo colonies by support of IL-5 as well as IL-3. Both CD34+ cells gradually lost the CD34 antigen by day 6 of incubation with G-CSF or IL- 3. Loss of this antigen was well-correlated with acquisition of susceptibility to IL-5. It was concluded that G-CSF supported the neutrophil differentiation of committed colony-forming cells, IL-3 supported that of both committed and multipotent colony-forming cells. G-CSF and IL-3 also supported the early stage of E. differentiation; IL- 5 supported the late stage of that.

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Expression of macrophage CD14 receptor in the course of experimental inflammatory responses induced by lipopolysaccharide and muramyl dipeptide

Veterinární Medicína ◽

10.17221/1991-vetmed ◽

2008 ◽

Vol 53 (No. 7) ◽

pp. 347-357 ◽

Cited By ~ 4

Author(s):

Z. Sladek ◽

D. Rysanek

Keyword(s):

Inflammatory Response ◽

Inflammatory Responses ◽

Early Stage ◽

Muramyl Dipeptide ◽

Cell Suspensions ◽

The Other ◽

Total Count ◽

The Difference ◽

Phosphate Buffered Saline ◽

Time Point

The aim of this study was to determine whether expression of CD14 on macrophages is regulated differently during initiation and resolution of the inflammatory response caused by CD14-dependent (lipopolysaccharide) and CD14-independent (muramyldipeptide) bacterial signals. In cell suspensions from the site of inflammation we observed two types of macrophages: non-vacuolized (NMAC) and vacuolized (VMAC) cells. NMAC (monocyte-like cells) were dominant during the early stage of the inflammatory response, whilst VMAC contained phagocytosed apoptotic neutrophils in various stages of digestion. These latter cells were dominant during resolution (particularly at the last time point of 168 h). Intramammary instillation of muramyldipeptide (MDP) and lipopolysaccharide (LPS) resulted in a significant increase in the total count of CD14+ NMAC after 24 h (muramyldipeptide P < 0.01 and lipopolysaccharide P < 0.05) compared to phosphate buffered saline (PBS). During resolution of the inflammatory response, a gradual decrease in the total count of CD14+ NMAC was observed. The difference compared with PBS was significant at 48 h and 72 h after instillation of both bacterial agents (muramyldipeptide: P < 0.05; lipopolysaccharide: P < 0.05). A lower total count of CD14+ VMAC was observed as an effect of MDP and LPS at 24 h after induction (P < 0.05), when compared to PBS. During resolution, the total count of CD14+ VMAC increased. Differences (P < 0.01) were observed at 72 h and 168 h after LPS compared to PBS. We therefore assume that the expression of CD14 on macrophages is not regulated differently during the inflammatory responses caused by CD14-dependent and CD14-independent bacterial signals. On the other hand, the stage of the inflammatory response to MDP and LPS played an important role in the regulation of CD14 expression on macrophages.

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THE EFFECT OF INORGANIC SALTS ON THE CHEMICAL CHANGES IN THE BLOOD OF THE DOG AFTER OBSTRUCTION OF THE DUODENUM

Journal of Experimental Medicine ◽

10.1084/jem.39.2.321 ◽

1924 ◽

Vol 39 (2) ◽

pp. 321-330 ◽

Cited By ~ 14

Author(s):

Russell L. Haden ◽

Thomas G. Orr

Keyword(s):

Sodium Chloride ◽

Inhibitory Action ◽

Sodium Citrate ◽

Fatal Outcome ◽

The Other ◽

Inorganic Salts ◽

Untreated Animal ◽

Protein Nitrogen ◽

Chemical Changes ◽

Therapeutic Value

The chemical changes in the blood of dogs treated with various inorganic salts after obstruction of the duodenum are reported. Two dogs treated with sodium chloride survived approximately six times as long as the average untreated animal, one living 22 days, the other 24 days. Ammonium chloride was found to produce an acidosis. The administration of potassium chloride, calcium chloride, and magnesium chloride did not prevent the usual rise in non-protein nitrogen and fall in chlorides, and the fatal outcome. Iodides seemingly hasten the toxic process. Sodium bromide appears to have an inhibitory action upon it, but much less than that of sodium chloride. Sodium sulfate, magnesium sulfate, sodium citrate, monosodium phosphate, and disodium phosphate failed to alter the course of the intoxication. Atropine and pilocarpine were without therapeutic value in preventing the changes characteristic of intestinal obstruction.

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Cytokine signaling during myocardial infarction: sequential appearance of IL-1 beta and IL-6

AJP Regulatory Integrative and Comparative Physiology ◽

10.1152/ajpregu.1995.269.2.r229 ◽

1995 ◽

Vol 269 (2) ◽

pp. R229-R235 ◽

Cited By ~ 17

Author(s):

I. Guillen ◽

M. Blanes ◽

M. J. Gomez-Lechon ◽

J. V. Castell

Keyword(s):

Myocardial Infarction ◽

Endothelial Cells ◽

Protein Synthesis ◽

Proinflammatory Cytokines ◽

Early Stage ◽

Myocardial Infarct ◽

Cytokine Signaling ◽

Target Cells ◽

Necrosis Factor Alpha

The purpose of this study was to investigate the significance of the sequential changes in proinflammatory cytokines observed in the plasma of patients early after myocardial infarct: a rise in interleukin (IL)-1 beta (308 +/- 126 vs. 141 +/- 78 pg/ml, P < 0.01) between 0 and 2 h followed by an IL-6 peak (49 +/- 24 vs. 14.5 +/- 13 pg/ml, P < 0.01) 4-9 h later. No significant changes in tumor necrosis factor-alpha (TNF-alpha) were observed at this early stage. The linkage between IL-1 beta and IL-6 secretions is supported by 1) the ability of patient's plasma drawn early after myocardial infarction to induce IL-6 mRNA and protein synthesis in cells that may be potential targets in vivo (fibroblasts and endothelial cells), 2) suppression of this activity by antibodies against IL-1 beta, and 3) a delay between IL-1 beta and IL-6 peaks in vivo (4-9 h), which is similar to the time required for maximal IL-6 production in IL-1 beta stimulated target cells in vitro (6 h). This sequential signaling might serve as the basis for an amplification mechanism of proinflammatory cytokines. In fact, a much greater synthesis of C-reactive protein was observed in hepatocytes when stimulated with conditioned medium of fibroblasts or endothelial cells that had previously been incubated with plasma of patients. The results of our work strongly suggest that, by inducing IL-6 in potential target cells, IL-1 beta could act as the primary, but indirect, signal that stimulates acute-phase protein synthesis after myocardial injury.

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STUDIES UPON EXPERIMENTAL MEASLES

Journal of Experimental Medicine ◽

10.1084/jem.36.2.231 ◽

1922 ◽

Vol 36 (2) ◽

pp. 231-238 ◽

Cited By ~ 4

Author(s):

Charles W. Duval ◽

Rigney D'Aunoy

Keyword(s):

Incubation Period ◽

Temperature Rise ◽

Early Stage ◽

Clinical Syndrome ◽

Human Infection ◽

Direct Result ◽

Cutaneous Manifestations ◽

The Face ◽

Specific Virus ◽

Repeated Passage

An active transmissible virus exists in the blood of measles patients during the eruptive stage of the disease. This virus produces in rabbits after intravenous injection a specific reaction analogous in all essential features to that of the human infection. Following a definite incubation period of from 2 to 5 days the animals infected show pyrexial, leucocytic, and cutaneous alterations. Fully 90 per cent of such inoculated rabbits react in a remarkable manner. The earliest constant symptom of the infection is a rise in temperature, which on the average occurs 4 days after inoculation and most probably marks the end of the incubation period. Concomitantly with this temperature rise there is a diminution in the total number of circulating leucocytes. This decrease in the number of white blood elements may be relative or may appear in the form of a well defined leucopenia. The most striking objective signs are the coryza, conjunctival injection, enanthemata, and exanthemata. The mucous membrane lesions are similar in their physical appearance to the so called Koplik spots seen in man. They occur on the buccal side of the oral cavity ranging in number from two to eight discrete hemorrhagic areas with paler centers. They appear as a rule coincidently with the temperature rise or shortly thereafter. The exanthematous lesions though occurring only in about 40 per cent of the infected animals complete the clinical syndrome in this particular experimental host. The rash may appear as early as the 3rd and as late as the 7th day after inoculation. In its early stage it is of the macular variety, appearing as a diffuse eruption which later develops into a more papular type of lesion. At this time the cutaneous manifestations appear as slightly raised, flattened, purplish red, discrete areas in the skin of the face, neck, chest, and abdomen. Repeated passage of the virus of measles through the rabbit seems to increase its virulence. A number of animals infected with such passage virus succumb in the fourth and subsequent generations, undoubtedly as the direct result of the action of the specific excitant, as in none of the animals was there cultural evidence of secondary intercurrent infection. In the animals dying presumably as a result of the specific virus grave nephritic changes were evident. It is a noteworthy fact that the pneumonia so common in fatal cases of human measles was not evident in any of the experimental animals. We believe this to be of considerable significance, especially in elucidating the direct etiological factor of the fatal pneumonias so often present in human measles cases. Apparently such infections in man can be explained purely on the basis of the destruction of normal defense barriers by the specific excitant of the infectious disease, and the lack of host resistance to the ordinary pyogenic microorganism.

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Different Expressions of IL-1 TNF-α P-selectin mRNA by Endothelial Cells after Vein Thrombosis

10.1101/681387 ◽

2019 ◽

Author(s):

Jiasheng Xu ◽

Kaili Liao ◽

Weimin Zhou

Keyword(s):

Endothelial Cells ◽

Early Stage ◽

Early Period ◽

Target Cells ◽

Mrna Levels ◽

Vein Thrombosis ◽

Tnf Α ◽

Group A ◽

Group B ◽

Porcine Endothelial Cells

[Abstract]ObjectiveExperiments were designed to compare the expressions of IL-1 TNF-α P-selectin mRNA by porcine endothelial cells after vein thrombosis.MethodsIVC under the renal vein of 20 porcines were ligated to induce thrombosis modes. These thrombosed veins were divided into three groups:group A-one day after thrombosis, group B-four days after thrombosis and group C-seven days after thrombosis. The other pigs were given the shame operation as a contro group (group D). The mRNA levels of IL-1、 TNF-α and P-selectin expressed by porcine endothelial cells in three groups were analy sed by semi quantitative RT-PCR. Endothelial cells were harvested with collagenase II.ResultsThe purity of endothelial cells harvested was 99.42 ±0.07. The expression of IL-1 was detained only in group A while TNF-αreached its peak in group B(P<0.05) and P-selectin increased gradually with the days passing by(P<0.05).ConclusionEndothelial cells are not only the target cells of inflammatory mediators, but also can express a variety of active factors to promote venous thrombosis. Expression of TNF-α mRNA is increased gradually in the early period of vein thrombosis whileP-selectin in the acute period; IL-1 mRNA was transiently expressed only in the early stage of thrombosis.

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