中枢去甲肾上腺素与E－2078和盐酸二氢埃托啡在痛觉调制中的相互作用

给大鼠脊髓蛛网膜下腔（ｉ．ｔ．）单独注射去甲肾上腺素（ＮＥ）２．μｇ或强啡肽Ａ（１～８）衍生物（Ｋ激动剂）Ｅ－２０７８８０ｎｇ可产生轻度但显著的镇痛效应。同时ｉ．ｔ．注射１／２量ＮＥ和１／２量Ｅ－２０７８所产生的镇痛效应显著高于单独ＮＥ或Ｅ－２０７８的效应。将Ｅ－２０７８的剂量增加到９６和１１５ｎｇ（依次递增２０％），也得到与上述类似的结果，提示Ｅ－２０７８与ＮＥ在脊髓产生协同镇痛作用。用同样的方法观察皮下注射（ｓ．ｃ．）μ激动剂盐酸二氢埃托啡（ＤＨＥ）对ｉ．ｔ．注射ＮＥ镇痛的影响，发现ｓ．ｃ．ＤＨＥ在２６８，３７５，５２５ｎｇ三个剂量水平（依次递增４０％）下，１／２量ＮＥ和１／２量ＤＨＥ联合注药组产生的镇痛效应与单纯ＮＥ或ＤＨＥ组相比均无显著性差异。以上结果提示，脊髓内的ＮＥ与。阿片激动剂Ｅ－２０７８有协同镇痛作用，而与μ阿片激动剂ＤＨＥ仅有加和作用。     

INDUCTION AND REVERSAL OF IMMUNE PARALYSIS IN VITRO

Induction of the immune response can only be completed after antigen is removed from the cellular environment. Primed rabbit lymph node fragments were cultured in vitro with 5 mg/ml BSA. If antigen was removed from the fragments 2 hr later, they produced a normal anti-BSA response, which was first evident 5 days later. If antigen removal was delayed for 3 days, the onset of the response was postponed for 2 to 3 days. Pulses with BUDR marked the periods of cell proliferation in both sets of cultures, and established that the postponement of antibody production was preceded by a postponement in the wave of proliferation among precursors of antibody forming cells. The similarity in avidity of antibody-containing fluids from normal and postponed cultures support the idea that the same cell population produced the response in each case. It was concluded that a reversible state of paralysis could be instituted in antigen-responsive cells, and this state did not depend upon cell-killing. The widespread incidence of temporary paralysis as an early aspect of the immune response was discussed.     

FATE OF ANTIGEN-BINDING CELLS IN UNRESPONSIVE AND IMMUNE MICE

Antigen-binding cells (ABC) to the antigen human gamma globulin (HGG) were quantitated in lymphoid tissues of A/J mice at various times after the injection of deaggregated HGG (tolerogen), aggregated HGG (immunogen), or saline. The reaction of lymphoid cells with highly labeled HGG was specific to that antigen since binding could be inhibited by excess unlabeled HGG, but not by unrelated non-cross-reacting proteins. Compared with normal mice, there was a marked decrease in the numbers of ABC in the spleens of unresponsive animals evident as early as 12 h after the injection of tolerogen. A marked increase in ABC was observed in the spleens of immunogen-injected mice, beginning at 24 h and reaching a peak at 3 days. In bone marrow, no difference in the number of ABC was found among the three experimental groups until day 20, when a reduction in ABC was observed only in tolerogen-injected mice. No quantitative difference in the thymuses in the experimental groups could be determined because of the paucity of ABC displayed by normal thymus cells.     

PASSIVE ANTIBODY AND THE IMMUNE RESPONSE : FACTORS WHICH DETERMINE ENHANCEMENT AND SUPPRESSION

The isoimmune response of fowl inoculated with RBC coated with antibody was investigated. Anti-B antiserum from a single animal was used to coat different donor type RBC. With each donor type RBC the immune response to the coated determinants is suppressed. Enhancement of the immune response to noncoated determinants occurs when they are products of an allelic gene or belong to a different blood group system. Coating some B antigen determinants suppresses the response to noncoated determinants of the same antigen, i.e., determinants which are products of the same B gene. Varying the quantity of passive antibody revealed that the degree of suppression and the degree of enhancement are negatively correlated. These findings support the concept that antibody-coated determinants function as carrier for noncoated determinants, provided a certain physical association exists between them. A further interpretation of these studies is that in certain situations an antibody to one antigen may interfere with events which lead to an immune response to a different antigen. The possibility, that the protection afforded by ABO incompatibility against Rh isoimmunization is because of a similar phenomenon, is discussed. A hypothesis is presented which states that where the immune response to certain antigens behaves as a dominantly inherited trait, and is associated with histocompatibility type, the nonresponder animals possess an antibody (perhaps cell bound) which interferes with the response to determinants for which it does not have specificity. Responders are assumed to lack this antibody because it has specificity for their major histocompatibility antigens.     

AN ELECTROPHORETIC STUDY OF IMMUNE SERA AND PURIFIED ANTIBODY PREPARATIONS

1. Antibody produced in the horse migrates as a new serum component between the β and γ components, whereas rabbit antibody is electrophoretically identical with the γ globulin component of the serum. 2. In rabbit and monkey antisera the percentage of antibody in the serum and in the γ globulin fraction can be determined by integration of the electrophoresis diagrams of unabsorbed and absorbed sera. Antibody solutions of high purity can be obtained by electrophoretic isolation of the γ globulin of rabbit antisera in which the percentage of antibody to total γ globulin is high. 3. The isoelectric points of pig, cow, horse, and rabbit antibodies have been determined. 4. In horse sera prolonged immunization is accompanied by the formation of another antibody component of lower mobility.     

IMMUNE AND NATURAL ANTIBODIES TO SYNGENEIC MURINE PLASMA CELL TUMORS

Cytotoxic antibody to a plasma cell tumor antigen was produced in syngeneic BALB mice by immunization with viable or inactivated plasma cell tumors. Antibody with the same specificity was found in the sera of normal BALB and other strains of mice. This natural antibody reacted with an antigen with characteristics indistinguishable from the previously described alloantigen, PC.1, and with viral envelope antigen, χVEA. The incidence of cytotoxic reactivity and the antibody titers reached a peak in normal BALB mice at 3–4 months of age, and were lower in 9–12-month old mice. The sera of germfree mice had lower reactivity; but when the mice were transferred to conventional conditions, their sera soon became as active as those of conventional mice. A virus common to all plasma cell tumors, which is present in latent form in some normal tissues of BALB and other PC.1 positive strains, is suggested as the cause for the PC.1 antigen and for the appearance of natural antibody to it. The considerable evidence for the close association of a virus with plasma cell tumors is presented.     

COMPONENTS OF GUINEA PIG COMPLEMENT : I. SEPARATION OF A SERUM FRACTION ESSENTIAL FOR IMMUNE HEMOLYSIS AND IMMUNE ADHERENCE

Employing sheep erythrocytes sensitized by antibody and the first and fourth components of complement (EAC'1,4), in such a manner as to prevent the development of immune adherence (I-A) reactivity during preparation, four separate substances required for the conversion of EAC'1,4,2 to the final damaged state (E*) were identified in whole guinea pig serum by cellulose chromatography, and tentatively termed C'3c, C'3b, C'3a, and C'3d. I-A reactivity was induced in EAC'1,4,2 after interaction with only one of these four substances, C'3c. A detailed comparison of the effects of heat, hydrazine, low pH, freezing, absorption by immune complexes, and elution from cellulose columns indicated that this same substance which was capable of imparting I-A reactivity to EAC'1,4,2 was also essential for immune hemolysis. Other experiments showed that I-A-reactive cells prepared either by treating EA with different concentrations of whole C' at 0°C, or by treating EAC'1,4,2 with C'3c, underwent lysis by C'2 + C'3b + a + d in proportion to the amount of whole C' or of C'3c used to make the cells reactive in I-A. These data provide strong evidence that a single factor, C'3c, is required both for the conversion of EAC'1,4,2 to an I-A-reactive complex (EAC'1,4,2,3c) and for the lysis of EAC'1,4,2 by C'3b + a + d. C'3c is the only one of the components studied which can induce I-A reactivity, and is the first to react with EAC'1,4,2. Formation of EAC'1,4,2,3c proceeds even at 0°C, but is much more rapid at elevated temperatures, showing a maximum in from 5 to 15 minutes at 37° or 30°C respectively. Prolonged incubation at these temperatures results in a decline in hemolytic reactivity without a noticeable effect on I-A. This loss was resolved into three phenomena: (a) a rapid loss of ability of SAC'1,4,2,3c to react with C'3b, presumably as a result of decay of the C'2 moiety in the complex, which is readily reversed by addition of fresh C'2; (b) a slow, irreversible spontaneous inactivation of SAC'1,4,2,3c; (c) a moderately rapid, irreversible inactivation of SAC'1,4,2,3c by some factor present in C'3c preparations.     

CELLS INVOLVED IN THE IMMUNE RESPONSE : IV. THE RESPONSE OF NORMAL AND IMMUNE RABBIT BONE MARROW AND LYMPHOID TISSUE LYMPHOCYTES TO ANTIGENS IN VITRO

Cell suspensions of immune rabbit lymph nodes and spleen were capable of undergoing blastogenesis and mitosis and of incorporating tritiated thymidine when maintained in culture with the specific antigen in vitro. They did not respond to other, non-cross-reacting antigens. The blastogenic response obtained with immune lymph node cells could be correlated with the antibody synthesizing capacity of fragment cultures prepared from the same lymph nodes. Cell suspensions of immune bone marrow responded to non-cross-reacting antigens only whereas cell suspensions of immune thymus, sacculus rotundus, and appendix did not respond when exposed to any of the antigens tested. On the other hand, neither fragments nor cell suspensions prepared from lymph nodes, spleen, and thymus of normal, unimmunized rabbits responded with antibody formation and blastogenesis when exposed to any of the antigens. However, normal bone marrow cells responded with marked blastogenesis and tritiated thymidine uptake. The specificity of this in vitro bone marrow response was demonstrated by the fact that the injection of a protein antigen in vivo resulted in the loss of reactivity by the marrow cell to that particular antigen but not to the other, non-cross-reacting antigens. Furthermore, bone marrow cells of tolerant rabbits failed to respond to the specific antigen in vitro. It was also demonstrated that normal bone marrow cells incubated with antigen are capable of forming antibody which could be detected by the fluorescent antibody technique. This response of the bone marrow cells has been localized to the lymphocyte-rich fraction of the bone marrow. It is concluded that the bone marrow lymphocyte, by virtue of its capacity to react with blastogenesis and mitosis and with antibody formation upon initial exposure to the antigen, a capacity not possessed by lymphocytes of the other lymphoid organs, has a preeminent role in the sequence of cellular events culminating in antibody formation.     

ANTIGENIC RELATIONSHIPS BETWEEN IMMUNE GLOBULINS AND CERTAIN RELATED PARAPROTEINS IN MAN

The antigenic properties of normal 19S γ-globulin, pathologic macroglobulins, β_2A-myeloma proteins, and Bence Jones proteins have been compared with 7S γ-globulin and the small 3.5S units derived from it by gel diffusion precipitin techniques. These studies demonstrate that the determinant groups on the 7S γ-globulin molecule responsible for the cross-reaction with each of the other proteins are associated with the two fragments of 7S γ-globulin which have the antibody-combining sites. The antigenic specificity of the 7S γ-globulin which distinguishes it from each of these proteins is associated primarily with the fragment that is richest in hexose and can not combine with antigen. However when compared with certain of the paraproteins additional antigenic specificity was also found to reside in the fragments with antibody-combining activity. The finding of similar antigenic relationships in rabbit γ-globulins suggests that some of the biological properties associated only with the 7S γ-globulins and not with the other immune globulins may reside in the fragment which also carries the antigenic specificity of the protein.     

THE NEUTRALIZATION OF ANTIPNEUMOCOCCUS IMMUNE BODIES BY INFECTED EXUDATES AND SERA

1. In empyema fluids resulting from infection with pneumococci there are present large amounts of soluble substances which have the property of neutralizing pneumococcus antibodies. 2. Similar substances are found in the blood of infected rabbits. 3. When immune serum is injected into infected rabbits the immune substances disappear very quickly, and therefore are prevented from activity in overcoming the infection. 4. When immune serum is administered to patients severely infected with pneumococci, the immune bodies may also disappear very rapidly, and this disappearance is probably associated with the presence of such soluble substances in the blood. 5. The serum only becomes effective when these substances are neutralized. 6. The study of agglutination curves is of value in showing why in certain instances favorable results have not followed the use of immune serum. 7. It is important that in severely injected patients the serum be administered early in the disease and that the initial dosage be large.     

IMMUNIZATION AGAINST VACCINIA BY NON-INFECTIVE MIXTURES OF VIRUS AND IMMUNE SERUM

1. Vaccine virus and specific immune serum mixed in such proportions as to produce no lesion when inoculated intradermally induce immunity when instilled into the nasal cavities of rabbits. 2. The mixture is also effective when inoculated subcutaneously. 3. Immunity is brought about with a minimum of systemic reaction and no local one. 4. When the mixtures are incompletely neutralized generalized vaccinia results.     

THYROIDECTOMY AND PARATHYROIDECTOMY WITH RELATION TO THE DEVELOPMENT OF IMMUNE SUBSTANCES

1. After thyroidectomy with partial parathyroidectomy the maximum and average hemolytic titers of the sera of rabbits injected intravenously with sheep blood are equal to or higher than those of normal animals similarly injected. 2. Thyroidectomy with partial parathyroidectomy does not inhibit antibody production. This fact is in accord with the results of Garibaldi, Launoy and Lévy-Bruhl, Lerda and Diez, and others. 3. Thyroidectomy with partial parathyroidectomy does not cause serious disturbance in the adult rabbit. If the operation is performed properly, the animals survive and only moderate cachexia develops in time. 4. After complete thyroparathyroidectomy a small proportion of the animals survive even after developing very severe tetany. Those that recover do not show further signs of serious disturbance, but in time develop a moderate degree of cachexia no greater than that of the thyroidectomized animals. 5. Thyroparathyroidectomized rabbits develop anti-sheep hemolysin of a uniformly low titer—on an average one-fifth that of the controls. 6. Injection of bovine blood into rabbits that survived complete thyroparathyroidectomy from 1 to 2 months previously results in the production of hemolysin of a uniformly low titer compared with that of normal animals similarly treated.     

ANTIGEN MODULATION OF THE IMMUNE RESPONSE : THE EFFECT OF DELAYED CHALLENGE ON THE AFFINITY OF ANTI-DINITROPHENYLATED BOVINE GAMMA GLOBULIN ANTIBODY PRODUCED IN ADOPTIVE RECIPIENTS

Secondary immune responsiveness to dinitrophenylated bovine gamma globulin (DNP-BGG) was transferred to heavily irradiated rats by means of small lymphocytes from the thoracic duct of immunized syngeneic donors. Affinity of the antibody produced by the adoptive recipients when challenged immediately was the same as that seen in immunized controls and cell donors, suggesting that the performance of lymphocytes in the thoracic duct of immunized rats provide an accurate measure of the immunologic history of memory cells both in the intact animal and the lymphocyte-depleted donor. The relevance of this to cell cooperation in the production of antibody to hapten-protein complexes is discussed. Direct evidence on antigen modulation of the immune response was also obtained. When immune thoracic duct lymphocytes were transferred to adoptive recipients and challenged either immediately or after a delay of 6 wk, it was found that, although the amount of antibody decreased with this delay in challenge, the affinity remained constant. Thus it would seem that in control rats, the increase in the affinity of antibody with time after immunization is, in fact, due to the selection of new populations of cells by the progressively waning concentration of antigen.     

IMMUNE RESPONSES IN VITRO : I. CELLULAR REQUIREMENTS FOR THE IMMUNE RESPONSE BY NONPRIMED AND PRIMED SPLEEN CELLS IN VITRO

A cell suspension culture system combined with a procedure which separates most macrophages from lymphoid cells was used to investigate some of the cellular requirements for direct and indirect plaque-forming cell responses by nonprimed and primed mouse spleen cells in vitro. The plaque-forming cell response to heterologous erythrocytes in cultures of nonprimed spleen cells required both macrophages and lymphoid cells for its development. A significant indirect plaque-forming cell response did not develop in cultures of nonprimed spleen cells. In contrast, cultures of separated or macrophage-poor lymphoid cells from primed mice exhibited increasing responses relative to the response of unseparated spleen cells as the interval after priming increased. The cultures of separated lymphoid cells were not entirely free of phagocytic cells. Despite some evidence which suggests that these phagocytic cells had little function in the response, one cannot ascertain whether the lymphoid cells were responding directly to a second contact with antigen or whether the few contaminating phagocytic cells were performing a function essential to the response by the lymphoid cells. Physiologically different populations of cells appear to develop after priming and are able to respond in vitro in a macrophage-poor culture. Some of the properties of these populations suggest that they are "memory cell" pools containing precursors of direct and indirect plaque-forming cells highly susceptible to a second antigenic stimulus.     

SPECIFIC IMMUNE RESPONSE GENES OF THE GUINEA PIG : V. INFLUENCE OF THE GA AND GT IMMUNE RESPONSE GENES ON THE SPECIFICITY OF CELLULAR AND HUMORAL IMMUNE RESPONSES TO A TERPOLYMER OFL-GLUTAMIC ACID,L-ALANINE,ANDL-TYROSINE

The ability of guinea pigs to make immune responses to the random linear copolymer of L-glutamic acid and L-alanine, GA, and to L-glutamic acid and L-tyrosine, GT, is each controlled by a different immune response gene. On the other hand, the random linear terpolymer of L-glutamic acid, L-alanine, and L-tyrosine, GAT, which contains both GA and GT antigenic determinants, is immunogenic in all guinea pigs. After GAT immunization, all animals develop delayed hvpersensitivity and serum antibody specific for GAT. However, only those guinea pigs possessing the GA immune response gene demonstrate cross-reactive delayed hypersensitivity when challenged with GA. In addition, the anti-GAT antisera produced by those animals having the GA gene contain cross-reacting anti-GA antibodies. The sera from guinea pigs lacking the GA gene have no anti-GA antibody activity. Thus, we have demonstrated that a specific immune response gene controlling responsiveness to a "simple" antigen can determine the specificity of both cellular and humoral immune responses to a more complex antigen.