Regional mapping of two human immunoglobulin V lambda genes and analysis of the V lambda locus in chronic myeloid leukaemia.

The human immunoglobulin V lambda locus has been studied in relation to chromosomal translocations involving chromosome 22. DNA probes for two V lambda genes which belong to different subgroups and do not cross hybridize, were used to show that both V lambda genes are located on the Philadelphia chromosome in chronic myeloid leukaemia (CML). Both genes map in band 22q11 to a region that is bounded on the distal side by the breakpoints for CML 9:22 translocations and on the proximal side by the breakpoint for an X:22 translocation. We have found no evidence for rearrangements or amplification of either V lambda gene in CML, in either the chronic or acute phases of the disease. In K562 cells which are derived from the pleural effusion of a patient with Ph1-positive CML, there appears to be no rearrangement of the V lambda genes, but they are both amplified about four times. We have estimated that the minimum size for the amplification unit in K562 cells is 186 kb.     

The V lambda J lambda repertoire in human fetal spleen: evidence for positive selection and extensive receptor editing

VlambdaJlambda rearrangements obtained from genomic DNA of individual IgM(+) B cells from human fetal spleen were analyzed. A nonrandom pattern of lambda gene rearrangements that differed from the adult Vlambda repertoire was found. The Vlambda distal genes 8A and 4B were absent from the nonproductive fetal repertoire, whereas 2E and 3L were overrepresented and 1B was underrepresented in the productive fetal repertoire. Positive selection of the Vlambda gene, 2E, along with Vlambda rearrangements employing homologous VlambdaJlambda joins were observed in the fetal, but not in the adult Vlambda repertoire. Overrepresentation of Jlambda distal cluster C genes rearranging to the Vlambda distal J segment, Jlambda7, in both productive and nonproductive fetal repertoires suggested that receptor editing/replacement was more active in the fetus than in adults. Numerous identical VlambdaJlambda junctions were observed in both the productive and nonproductive repertoire of the fetus and adult, but were significantly more frequent in the productive repertoire of the fetus, suggesting expansion of B cells expressing particular lambda-light chains in both stages of development, with more profound expansion in the fetal repertoire. Notably, B cells expressing identical lambda-light chains expressed diverse heavy chains. These data demonstrate that three mechanisms strongly influence the shaping of the human fetal lambda-chain repertoire that are less evident in the adult: positive selection, receptor editing, and expansion of B cells expressing specific lambda-light chains. These events imply that the expressed fetal repertoire is shaped by exposure to self Ags.     

Serologically defined V region subgroups of human lambda light chains

The availability of numerous antisera prepared against lambda-type Bence Jones proteins and lambda chains of known amino acid sequence has led to the differentiation and classification of human lambda light chains into one of five V lambda subgroups. The five serologically defined subgroups, V lambda I, V lambda II, V lambda III, V lambda IV, and V lambda VI, correspond to the chemical classification that is based on sequence homologies in the first framework region (FR1). Proteins designated by sequence as lambda V react with specific anti-lambda II antisera and are thus included in the V lambda II subgroup classification. The isotypic nature of the five V lambda subgroups was evidenced through analyses of lambda-type light chains that were isolated from the IgG of normal individuals. Based on analyses of 116 Bence Jones proteins, the frequency of distribution of the lambda I, lambda II/V, lambda III, lambda IV, and lambda VI proteins in the normal lambda chain population is estimated to be 27%, 37%, 23%, 3%, and 10%, respectively. This distribution of V lambda subgroups was comparable to that found among 82 monoclonal Ig lambda proteins. Considerable V lambda intragroup antigenic heterogeneity was also apparent. At least two sub-subgroups were identified among each of the five major V lambda subgroups, implying the existence of multiple genes in the human V lambda genome. The V lambda classification of 54 Ig lambda proteins obtained from patients with primary or multiple myeloma-associated amyloidosis substantiated the preferential association of lambda VI light chains with amyloidosis AL and the predominance of the normally rare V lambda VI subgroup in this disease.     

Immunogenic and antigenic epitopes of Ig. XXV. Monoclonal antibodies that differentiate the Mcg+/Mcg- and Oz+/Oz- C region isotypes of human lambda L chains

The C region of human lambda L chains is specified by multiple C lambda genes of which three--C lambda 1, C lambda 2, and C lambda 3--encode for the isotypes designated Mcg+, Kern- Oz-, and Kern- Oz+, respectively. The Mcg, Kern, and Oz factors have been characterized by sequence differences involving specific C lambda amino acid residues. They have also been recognized serologically by polyclonal antisera but, with rare exception, these reagents are no longer available. We have obtained two murine anti-human lambda-chain mAb, 14G1 and 14D1, that recognize antigenic determinants specific for the C lambda isotypes Mcg and Oz, respectively. These antisera have been used to classify as Mcg+/Mcg- or Oz+/Oz- monoclonal lambda-chains (Bence Jones proteins) and intact Ig lambda proteins. There was complete concordance between the chemical and serologic assignment of lambda-chains as Mcg+/Mcg- or as Oz+/Oz-; no single protein expressed both isotypes. There was no evident association between the C region isotype Mcg or Oz and the V region subgroup of the protein tested. However, our finding that four of seven amyloid-associated lambda VI Bence Jones proteins were Oz+ suggests a predominant expression of the C lambda 3 gene product among proteins of this uncommon V lambda subgroup.     

Wild mice express an Ig V lambda gene that differs from any V lambda in BALB/c but resembles a human V lambda subgroup.

The lambda L chain locus in the inbred mouse strains commonly used in the laboratory contains a limited number of germ-line genes; only three V lambda and three functional J lambda-C lambda genes have been identified in BALB/c mice. Previous studies indicated that wild mice may have a considerably expanded number of C lambda genes, as judged by the number of DNA restriction fragments that hybridize to C lambda probes derived from BALB/c. In order to evaluate the expression of these putative lambda genes, we have determined sequences of cDNA encoding lambda-chains in hybridomas from wild mice of the subspecies Mus musculus musculus from two different geographic regions, Denmark and Czechoslovakia. Two of these hybridomas produce L chains with J and C regions that are very similar to those of BALB/c lambda 1 chains, but the V regions of these L chains are only approximately 40% identical in amino acid sequence to the known murine V lambda. Indeed, these wild mouse V lambda are closer in sequence to human V lambda than they are to BALB/c V lambda, especially to human V lambda of subgroup VI, with which they share an unusual two-residue insertion in framework 3; L chains bearing V regions of this rare human type have a marked tendency to enter into amyloid deposits. These findings suggest that similar V lambda may be widespread in mammalian populations, although analysis by Southern blotting indicates that they are not found in BALB/c mice. A third hybridoma produces a L chain whose V lambda resembles BALB/c V lambda 1. The J lambda and C lambda segments of the cDNA encoding all three hybridoma L chains are identical; evidently, of the several putative genes that hybridize to C lambda 1 probes, one is expressed preferentially.     

Depression of the vestibulo-ocular and optokinetic responses by intrafloccular microinjection of GABA-A and GABA-B agonists in the rabbit

The functional implication of the cerebellar flocculus in regulation of the VOR and OKR gain has mostly been studied by lesion experiments, and the hypotheses derived from these experiments are not always in line with one another. In the present study, a reversible method was used to inhibit floccular Purkinje cells. The GABA-A agonist muscimol or the GABA-B agonist baclofen were bilaterally injected into the flocculus of rabbits, and the effects of these injections on the gain of the VOR and OKR were studied. Both drugs induced a reduction by at least 50% of the gain of the VOR in light and darkness, and of the OKR. Although GABA-A and GABA-B receptors are known to have different cerebellar localizations, muscimol and baclofen injections resulted in quantitatively similar effects. It is suggested that these GABA-agonists cause either direct or indirect inhibition of floccular Purkinje cells, thus reducing modulation of the firing rate of these neurons by afferent mossy and climbing fibers. Because the flocular Purkinje cells act out of phase with the vestibular neurons which drive the oculomotor neurons, a reduced output of floccular Purkinje cells would result in a reduction of the VOR and OKR gain. These experiments provide strong evidence that the cerebellar flocculus has a positive influence on the basic VOR and OKR gain.     

Analysis and Modeling of Frequency-Specific Habituation of the Goldfish Vestibulo-Ocular Reflex

Modification of the vestibulo-ocular reflex (VOR) by vestibular habituation is an important paradigm in the study of neural plasticity. The VOR is responsible for rotating the eyes to maintain the direction of gaze during head rotation. The response of the VOR to sinusoidal rotation is quantified by its gain (eye rotational velocity/head rotational velocity) and phase difference (eye velocity phase—inverted head velocity phase). The frequency response of the VOR in naïve animals has been previously modeled as a high-pass filter (HPF). A HPF passes signals above its corner frequency with gain 1 and phase 0 but decreases gain and increases phase lead (positive phase difference) as signal frequency decreases below its corner frequency. Modification of the VOR by habituation occurs after prolonged low-frequency rotation in the dark. Habituation causes a reduction in low-frequency VOR gain and has been simulated by increasing the corner frequency of the HPF model. This decreases gain not only at the habituating frequency but further decreases gain at all frequencies below the new corner frequency. It also causes phase lead to increase at all frequencies below the new corner frequency (up to some asymptotic value). We show that habituation of the goldfish VOR is not a broad frequency phenomena but is frequency specific. A decrease in VOR gain is produced primarily at the habituating frequency, and there is an increase in phase lead at nearby higher frequencies and a decrease in phase lead at nearby lower frequencies (phase crossover). Both the phase crossover and the frequency specific gain decrease make it impossible to simulate habituation of the VOR simply by increasing the corner frequency of the HPF model. The simplest way to simulate our data is to subtract the output of a band-pass filter (BPF) from the output of the HPF model of the naïve VOR. A BPF passes signals over a limited frequency range only. A BPF decreases gain and imparts a phase lag and lead, respectively, as frequency increases and decreases outside this range. Our model produces both the specific decrease in gain at the habituating frequency, and the phase crossover centered on the frequency of habituation. Our results suggest that VOR habituation may be similar to VOR adaptation (in which VOR modification is produced by visual-vestibular mismatch) in that both are frequency-specific phenomena.     

The immunoglobulin lambda locus in rat consists of two C lambda genes and a single V lambda gene

The immunoglobin lambda locus of the rat has been studied. Germ-line V lambda and C lambda genes were isolated from recombinant-phage libraries and characterized by nucleotide sequencing. The results showed that the lambda locus of the rat contains one single V lambda gene and two C lambda genes, thus representing one of the least complex lambda loci so far characterized. The two C lambda genes are separated by a spacer approx. 3 kb long. Two J segments are located at the 5' side of each C lambda gene. One of the C lambda genes (C lambda 1) probably represents a pseudogene, as the J lambda 1 segments have non-functional recombination and splice signals. The organization of the rat lambda locus resembles that of mouse, except that only one cluster is present in the rat. Thus since the evolutionary separation of the rat and mouse species ten MYR ( = 10(6) years) ago, either one cluster has been lost from the rat, or duplicated in the mouse.     

Transient analysis of vestibular nystagmus

The significance of a nystagmus-dependent, transient component in the overall slow-phase response of the vestibulo-ocular reflex (VOR) is brought into focus. First, a simulated example is presented that shows how this transient component can bias current algorithms for the estimation of VOR parameters. Second, new algorithms are proposed that are able to estimate VOR parameters regardless of the presence of transients. Third, the new algorithms are applied to experimental data, and the results are compared with those from current algorithms. The results clearly show that the transient component can significantly alter the apparent VOR time constant, particularly when the reflex has been lesioned. The algorithms open new areas of research on the possible role of nystagmus in enhancing the compensatory function of the VOR.     

Molecular characterization of a human anti-Rh(D) antibody with a DH segment encoded by a germ-line sequence.

The lambda-light-chain and lambda-heavy-chain variable-region genes of an anti-Rh(D) (Rh, Rhesus; D, heavy-chain diversity region) human monoclonal antibody secreted by lymphocytes transformed by the Epstein-Barr virus have been cloned and sequenced. Sequence comparison of the anti-Rh(D)mAb lambda-chain variable region with those of the other available human lambda chains revealed that it belonged to the human V lambda I (V lambda, variable region of lambda chain) subgroup. The greatest sequence similarity (80%) was observed with that of another anti-Rh antibody lambda-chain directed against the Rh(c) antigen. For the VH (VH, variable region of heavy chain) sequence, the highest similarity (86%) was observed with the germline VHG3 gene which belongs to the VHI subgroup. The expressed DH sequence of the anti-Rh(D) antibody is also of germline origin and complementarity-determining region 3 is thus produced by VH-DH and DH-JH (J, joining region) joining without recombination of multiple DH gene segments.     

Characterization of the human Ig V lambda II gene family and analysis of V lambda II and C lambda polymorphism in systemic lupus erythematosus.

We report the cDNA sequence of an expressed human V lambda II gene and present an RFLP analysis of the Ig gene family defined by this clone. This V lambda II gene was expressed in a monoclonal B cell line generated from a patient with SLE by transformation with EBV. The encoded lambda L chain displays the 8.12 Id, an Id common to anti-DNA antibodies from patients with SLE. Using a coding region probe we estimate from Southern blot analysis that the germline V lambda II gene family contains at least 15 members. Many of the V lambda II restriction fragments are polymorphic both in SLE patients and in nonautoimmune individuals. EcoRI, HindIII, and TaqI RFLP analyses of the V lambda II gene family and EcoRI analysis of the C lambda gene family reveal no polymorphisms specific to SLE. Observed V lambda II and C lambda allele frequencies are the same among SLE patients and nonautoimmune individuals, and show no evidence of linkage disequilibrium between the two loci.     

New subgroups in the human T cell rearranging V gamma gene locus.

Two new V gamma genes in humans are described from rearrangement in T cell lines, which constitute single members of new V gene subgroups of the T-cell rearranging gamma (TRG gamma) locus. These two genes (herein designated as belonging to V gamma III and V gamma IV subgroups) are located between V gamma I/V gamma II subgroups and the constant (C) gamma genes. The existence of these new genes brings the number of different, potentially useable, human TRG V gamma genes to eight (excluding at least five pseudo V gamma genes) and the number of distinct subgroups to four. Polymorphism in the sequence of the V gamma II subgroup gene is also described and rearranged fragment sizes which make possible an unequivocal assignment of a V gamma rearrangement are given. These results extend previous conclusions of the inherited diversity of the human TRG V gamma locus.     

Variation in V lambda genes in the genus Mus

The complement of Ig V lambda genes in nine species of feral mice representing the four extant subgenera of the genus Mus was examined and compared with that of BALB/c inbred mice. Although all inbred strains examined have two V lambda genes, there is variation in the number of copies of V lambda genes in the wild mice. All feral representatives of M. musculus domesticus, from which inbred strains are derived, have at least three V lambda genes, indicating that a V lambda gene may have been lost during the inbreeding process. At least three V lambda genes are also found in representatives of three other M. musculus subspecies, including the stock of M. musculus musculus "Czech II" shown to have at least 12 C lambda genes. In comparing the complement of V lambda and C lambda genes in these animals, evidence is found that supports a mechanism of lambda gene reiteration involving duplication of a unit containing a V lambda and two C lambda genes. However, the possibility that C lambda gene amplification occurred independent of V lambda gene evolution cannot be ruled out. M. spicelegus and M. spretus, species that are semifertile with M. musculus, have one to three V lambda genes. Species more distantly related to M. musculus, such as M. cookii and M. platythrix, appear to have more (four to six) V lambda genes. Greater V lambda gene heterogeneity is also found in these animals. We propose that the ancestors of the subgenus Mus had more V lambda genes than are seen in modern species and that the paucity of V lambda genes in M. musculus, M. spicelegus, and M. spretus may be the result of V lambda gene deletion events that occurred since the divergence of the ancestor of these three species and those of the distantly related species.     

Control and modulation of canal driven vestibulo-ocular reflex.

It has been well known that the canal driven vestibulo-ocular reflex (VOR) is controlled and modulated through the central nervous system by external sensory information (e.g. visual, otolithic and somatosensory inputs) and by mental conditions. Because the origin of retinal image motion exists both in the subjects (eye, head and body motions) and in the external world (object motion), the head motion should be canceled and/or the object should be followed by smooth eye movements. Human has developed a lot of central nervous mechanisms for smooth eye movements (e.g. VOR, optokinetic reflex and smooth pursuit eye movements). These mechanisms are thought to work for the purpose of better seeing. Distinct mechanism will work in appropriate self motion and/or object motion. As the results, whole mechanisms are controlled in a purpose-directed manner. This can be achieved by a self-organizing holistic system. Holistic system is very useful for understanding human oculomotor behavior.     

Implications of gain modulation in brainstem circuits: VOR control system

Gain modulation is believed to be a common integration mechanism employed by neurons to combine information from various sources. Although gain fields have been shown to exist in some cortical and subcortical areas of the brain, their existence has not been explored in the brainstem. In the present modeling study, we develop a physiologically relevant simplified model for the angular vestibulo-ocular reflex (VOR) to show that gain modulation could also be the underlying mechanism that modifies VOR function with sensorimotor context (i.e. concurrent eye positions and stimulus intensity). The resulting nonlinear model is further extended to generate both slow and quick phases of the VOR. Through simulation of the hybrid nonlinear model we show that disconjugate eye movements during the VOR are an inevitable consequence of the existence of such gain fields in the bilateral VOR pathway. Finally, we will explore the properties of the predicted disconjugate component. We will demonstrate that the apparent phase characteristics of the disconjugate response vary with the concurrent conjugate component.     

Inactivation of bacteriophage lambda by combined X-ray and U.V.-light exposure.

Extracellular phage lambda has been successively exposed to X-rays and U.V. light. The plaque-forming ability of the irradiated phages was determined on host cells with different repair capacities. No change in sensitivity was found with a pre-treatment of one type of radiation to lethal damage inflicted by the other. This indicates that a prerequisite for an interaction of different types of radiation is either an active metabolism or repair process occurring during the two radiation exposures.     

Afferent signals from the extraocular muscles of the pigeon modify the electromyogram of these muscles during the vestibulo-ocular reflex.

There is no general agreement on whether afferent signals from the extraocular muscles play any part in oculomotor control. However, we have previously shown that they modify the responses of cells in the oculomotor control system during the vestibulo-ocular reflex (VOR). If, as we suspect, these signals have an important role in the control of the VOR from moment-to-moment, we should be able to demonstrate similar, functionally significant, modifications at the output of the reflex. We have recorded the electromyographic activity of several extraocular muscles of the right eye during the VOR and while imposing movements on the left eye. We describe how the activity of the muscles, reflected in the electromyogram, is modified in specific ways depending on the parameters of the imposed eye movements. The effects of the extraocular afferent signals on the eye-muscle responses to vestibular drive during the slow phase of the VOR appear to be corrective. Thus the present results provide strong evidence that afferent signals from the extraocular muscles are concerned in the control of the reflex from moment-to-moment, and suggest that the wider question of their role in oculomotor control merits further consideration.