Specificity of Passively Transferred Delayed Hypersensitivity

Najarían, John S.; Feldman, Joseph D.

doi:10.1084/jem.118.3.341

Cited by 70 publications

(20 citation statements)

References 5 publications

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“…Also, peril)heral blood cells are capable of producing an in vitro denvyelinating factor, possibly antibody before the onset of EAE symptoms (17). The low number of specifically-reactive cells demonstrated in the target organ in the present study is consistent with isotopic studies on passive transfer in delayed hypersensitivity, where the donor cells were found to comprise less than 4% of the infiltrating cells and to resemble large lymphocytes (18,19). Over 90% of the infiltrating cells belonged to the bone marrow-macrophage line and originated from the recipient.…”

Section: Discussionsupporting

confidence: 87%

Specific Binding of Peroxidase-Labeled Myelin Basic Protein in Allergic Encephalomyelitis

Johnson

Wı́sniewski

Raine

et al. 1971

Proc. Natl. Acad. Sci. U.S.A.

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A conjugate of horseradish peroxidase and the encephalitogenic basic protein from myelin has been used to study the antigen reactivity of tissue in the autoimmune disease, experimental allergic encephalomyelitis. Control conjugates were also prepared of peroxidase and bovine serum albumin and of peroxidase and lysozyme, another basic protein. The basic protein from myelin conjugate was specifically bound by lymph node cells from rabbits immunized against the basic protein.Some of these cells appeared to be plasma cells. The conjugate was also specifically bound by occasional cells in the spinal-cord infiltrates of animals with early signs of allergic encephalomyelitis. These cells resembled large lymphocytes and plasma cells. There was no difference between the binding of basic protein of bovine and rabbit origin. The findings suggest the possibility that a local release of antibody within the target organ may play a role in the pat4ogenesis of allergic encephalomyelitis.Experimental allergic encephalomyelitis (EAE) is an autoimmune disease believed to be a manifestation of delayed hypersensitivity to the encephalitogenic basic protein of myelin (BP) (1-4). However, the pathogenetic mechanisms involved in EAE, and whether antibody participates in the disease process, are far from clear. BP was coupled to peroxidase with glutaraldehyde, which reacts with the e-amino groups of lysine (7), by the procedure developed by Avrameas (5) for other proteins. It was necessary to use small quantities of BP, which has many lysine groups, in order to obtain a soluble product. The molecular ratio of protein to peroxidase of about 1 to 7, however, was similar to that recommended by Avrameas. The conjugate was prepared by dissolving 0.3-0.6 mg of BP and 6 mg of peroxidase (type VI, RZ 3.1; Sigma) in 0.4 ml of 0.1 M phosphate buffer (pH 6.8). Then, 0.1 ml of 0.25% glutaraldehyde, diluted with phosphate buffer from an 8% aqueous preparation (Polysciences), was added dropwise with gentle mixing. After the mixture had stood for 2 hr at room temperature, the unbound glutaraldehyde was removed by dialysis for 24 hr at 40C against three 2-liter changes of phosphate-buffered saline (PBS; 0.01 M phosphate buffer (pH 7.2)-0.85% NaCl). Centrifugation at 36,000 X g for 30 min at 40C removed any insoluble product that formed. Several mnodificationss of the Avrameas procedure were tried, such as changing the concentration of peroxidase or glutaraldehyde, the pH and the conjugation time, or adding BP to peroxidase previously treated with glutaraldehyde. These modifications often gave active conjugates, but did not offer any advantages. The final BP-peroxidase conjugate was soluble and remained active for more than 2 months when stored frozen in small aliquots to minimize multiple freezings and thawings. As controls, conjugates of peroxidase *with bovine serum albumin (BSA) (Sigma) and with lysozyme from egg white (Sigma) were prepared identically, except that 2.5 mg and 0.4 mg, respectively, were used. Lysozyme is a protein similar to ...

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Section: Discussionsupporting

confidence: 87%

Specific Binding of Peroxidase-Labeled Myelin Basic Protein in Allergic Encephalomyelitis

Johnson

Wı́sniewski

Raine

et al. 1971

Proc. Natl. Acad. Sci. U.S.A.

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“…Early studies based on adoptive transfer of radiolabeled primed lymph node cells led to the conclusion that the majority of the lymphocytes in the brain are not specific for BP (4) and that the blood-brain barrier is permeable to lymphocytes without respect to their antigen specificity (5). Yet other studies pointed to a preferential accumulation of specific T cells in the lesion (2,6). In the one study that examined directly the frequency of BP-responsive cells in the spinal cord, a similar frequency was found in the primed popliteal lymph nodes (7).…”

mentioning

confidence: 80%

“…In the last 30 years several groups of investigators have attempted to study the nature and the specificity of the autoimmune infiltrate in EAE (4)(5)(6)(7)(8). Early studies based on adoptive transfer of radiolabeled primed lymph node cells led to the conclusion that the majority of the lymphocytes in the brain are not specific for BP (4) and that the blood-brain barrier is permeable to lymphocytes without respect to their antigen specificity (5).…”

mentioning

confidence: 99%

T cells in the lesion of experimental autoimmune encephalomyelitis. Enrichment for reactivities to myelin basic protein and to heat shock proteins.

Mor

Cohen²

1992

J. Clin. Invest.

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To characterize the cellular immune response in an autoimmune lesion, we investigated the accumulation of specific T cells in the central nervous system in actively induced experimental autoimmune encephalomyelitis (EAE) In the last 30 years several groups of investigators have attempted to study the nature and the specificity of the autoimmune infiltrate in EAE (4-8). Early studies based on adoptive transfer of radiolabeled primed lymph node cells led to the conclusion that the majority of the lymphocytes in the brain are not specific for BP (4) and that the blood-brain barrier is permeable to lymphocytes without respect to their antigen specificity (5). Yet other studies pointed to a preferential accumulation of specific T cells in the lesion (2, 6). In the one study that examined directly the frequency of BP-responsive cells in the spinal cord, a similar frequency was found in the primed popliteal lymph nodes (7). Recently, Hickey et al. (8) analyzed the kinetics ofcell migration into the CNS after intravenous inoculation of encephalitogenic and control cells (8). These authors concluded that activated T cells irrespective ofantigen specificity penetrate the CNS within hours of injection, however, only BP-specific cells are retained in the nervous tissue 72 h after inoculation (8).We conducted the present study to examine the question of the frequency of BP-responsive T cells in the EAE lesion. We also investigated whether the EAE lesion might be enriched in T cells responsive to heat shock protein (hsp) antigens. Hsp65 is interesting because it is a dominant antigen in the Mycobacteria present in the CFA and because T cells responsive to hsp65 have been found to function in autoimmune arthritis (9) and in autoimmune diabetes (10, 1 1). We also studied alloreactive T cells because such T cells are not known to cause autoimmune disease and they are detectable in high frequency without the need for priming.The results of this study indicate that the CNS infiltrate is highly enriched for encephalitogenic anti-BP T cells but also accumulates T cells reactive to hsp65. Alloreactive T cells, in contrast, were present in the lesions at the same frequency as in the lymph nodes. The frequency of BP-reactive T cells was highest at the peak of paralysis; but these T cells also could be found in the lesion at lower frequencies long after clinical recovery. Some T cell lines recovered from the autoimmune infiltrate were found to be encephalitogenic. MethodsRats. Inbred Lewis rats were supplied by the Animal Breeding Center of this Institute and were used at 2-3 mo of age. Rats were matched for age and sex in each experiment.

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“…Only a few sensitized cells are required to initiate a delayed hypersensitivity response (33,34). The expression of the reaction requires cells of the monocyte-macrophage series (35).…”

Section: Discussionmentioning

confidence: 99%

Effects of cytotoxic immunosuppressants on tuberculin-sensitive lymphocytes in guinea pigs.

Winkelstein¹

1975

J. Clin. Invest.

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Specificity of Passively Transferred Delayed Hypersensitivity

Cited by 70 publications

References 5 publications

Specific Binding of Peroxidase-Labeled Myelin Basic Protein in Allergic Encephalomyelitis

Specific Binding of Peroxidase-Labeled Myelin Basic Protein in Allergic Encephalomyelitis

T cells in the lesion of experimental autoimmune encephalomyelitis. Enrichment for reactivities to myelin basic protein and to heat shock proteins.

Effects of cytotoxic immunosuppressants on tuberculin-sensitive lymphocytes in guinea pigs.

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